lea!!!] Effects Research Series
DIETARY  SUBACUTE TOXICITY OF ETHYLENE
         THIOUREA  IN THE LABORATORY RAT
                               Health Effects Research Laboratory
                              Office of Research and Development
                              U.S. Environmental Protection Agency
                        Research Triangle Park, North Carolina 27711

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                RESEARCH REPORTING SERIES

Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into nine series. These nine broad cate-
gories were established to facilitate further development and application of en-
vironmental technology.  Elimination of traditional grouping  was consciously
planned to foster technology transfer and a maximum interface in related fields.
The nine series are:

      1.  Environmental  Health Effects Research
      2.  Environmental  Protection Technology
      3.  Ecological Research
      4.  Environmental  Monitoring
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This report has been assigned to the ENVIRONMENTAL HEALTH EFFECTS RE-
SEARCH series. This series describes projects and studies relating to the toler-
ances of man for unhealthful substances or conditions. This work is generally
assessed from a medical viewpoint, including physiological or psychological
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clude biomedical instrumentation and health research techniques utilizing ani-
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This document is available to the public through the National Technical Informa-
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                                   EPA-600/1-77-023
                                   May  1977
  DIETARY SUBACUTE TOXICITY OF
    ETHYLENE THIOUREA IN  THE
         LABORATORY RAT
                by

      Ralph I.  Freudenthal
 Battelle Columbus Laboratories
         505 King Avenue
      Columbus, Ohio 43201
     Contract No.  68-02-1715
         Project  Officer

          Ronald L.  Baron
 Environmental Toxicology Division
 Health Effects Research Laboratory
 Research Triangle Park, N.C. 27711
U.S. ENVIRONMENTAL PROTECTION AGENCY
 OFFICE OF RESEARCH AND DEVELOPMENT
 HEALTH EFFECTS RESEARCH LABORATORY
 RESEARCH TRIANGLE PARK, N.C. 27711

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                           DISCLAIMER

     This report has been reviewed by the Health Effects Research
Laboratory, U.S. Environmental Protection Agency, and approved for
publication.  Approval does not signify that the contents necessarily
reflect the views and policies of the U.S. Environmental Protection
Agency, nor does mention of trade names or commercial products
constitute endorsement or recommendation for use.
                                 ii

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                               FOREWORD

     The many benefits of our modern, developing, industrial society are
accompanied by certain hazards.  Careful assessment of the relative risk
of existing and new man-made environmental hazards is necessary for the
establishment of sound regulatory policy.  These regulations serve to
enhance the quality of our environment in order to promote the public
health and welfare and the productive capacity of our Nation's population.

     The Health Effects Research Laboratory, Research Triangle Park,
conducts a coordinated environmental health research program in toxicology,
epidemiology, and clinical studies using human volunteer subjects.  These
studies address problems in air pollution, non-ionizing radiation,
environmental carcinogenesis and the toxicology of pesticides as well as
other chemical pollutants.  The Laboratory develops and revises air quality
criteria documents on pollutants for which national ambient air quality
standards exist or are proposed, provides the data for registration of new
pesticides or proposed suspension of those already in use, conducts research
on hazardous and toxic materials, and is preparing the health basis for
non-ionizing radiation standards.  Direct support to the regulatory function
of the Agency is provided in the form of expert testimony and preparation of
affidavits as well as expert advice to the Administrator to assure the
adequacy of health care and surveillance of persons having suffered imminent
and substantial endangerment of their health.

     The significance of the occurrence of residues of pesticides or
metabolites of pesticides in or on plants destined for human consumption
must be considered with respect to its ultimate impact on human health.
The present study evaluates the effect of the presence of ethylenethiourea
(ETU) in the diet of a mammalian model test system.  ETU is the major
metabolite of a class of fungicides (ethylenebisdithiocarbamates) and the
toxicological hazard associated with this biologically active metabolite
as a residue in food is under intensive investigation.  An evaluation of
the health hazards associated with the presence of ETU in food includes
the primary effects on the thyroid and liver as well as several associated
biochemical parameters in an effort to determine a dietary level which
might be safe for man.
                                          John H. Knelson, M.D.
                                               Director,
                                   Health Effects Research Laboratory
                                   iii

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                                ABSTRACT
     Ethylene thiourea (ETU) was fed to groups of rats at 0, 1, 5, 25,
125 or 625 ppm for up to 90 days.  Other groups of rats received either
propylthiouracil (PTU; 125 ppm) or amitrole (50 ppm) in their diets as
positive controls.  Only those rats which received ETU at 125 or 625 ppm
and those ingesting PTU or amitrole demonstrated a measurable toxic
response.  This toxicity was reflected as an alteration in thyroid func-
tion and a significant change in thyroid morphology.

     Ingestion of 625 ppm ETU, or 125 ppm PTU resulted in very substantial
decreases in serum triiodothyronine (T-3) and thyroxine (T-4).  Marked
increases in serum thyroid stimulating hormone (TSH) levels were found
in the 625 and 125 ppm ETU rats, the 125 PTU rats and the rats receiving
amitrole, at each time point this hormone was measured.  A decrease in
iodide uptake by the thyroid was also found in the rats which ingested
625 ppm ETU.  While a statistically significant increase in serum T-4
and degree of thyroid hyp.erpla.sia was observed for the rats ingesting
25 ppm ETU for 60 days, normal thyroid hormone levels and thyroid mor-
phology was found in the rats on 25 ppm ETU for either 30 or 90 days.

     Bas_.ed on. biochemical and microscopic changes examined, the no-effect
level for dietary ETU ia this 90rday study is considered to be 25 ppm,
equivalent to an average intake ranging from 19.5 mg/kg body weight at
Week 1 to 12.5 mg/kg body weight at Week 12.
                                      iv

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                               CONTENTS

Foreword	iii
Abstract	iv
Figures	vi
Tables	vii

     Introduction	   1
     Experimental	   2
          Materials	   2
          ETU Purity Analysis	   2
          Thyroid Function and Control 	   2
          Evaluation of Test Kits	   3
          Methods	   4
     Results 	   5
     References. .. -.	   9

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                                FIGURES

Number                                                               Page
  1   MS Analysis of the ETU used as the test chemical	    11
  2   MS of the ETU used as the test chemical	    12
  3   Diet consumption	    13
  4   Test chemical consumption 	    14
  5   Body weight gain	    15
                                     vi

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                                TABLES
Number                                                                Page
   1   ETU Protocol Design	   16
   2   Tissues Removed and Weighed During Necropsy 	   16
   3   Thyroid Hormone Levels - After 30 Days on Study 	   17
   4   Thyroid Hormone Levels - After 60 Days on Study 	   18
   5   Thyroid Hormone Levels - After 90 Days on Study 	   19
   6   Free Thyroxine Index	   20
   7   Weekly Diet Consumption 	   21
   8   Weekly Test Chemical Ingestion	   23
   9   Weekly Body Weight Gain	   24
  10   Organ Weights Obtained at Necropsy	   26
  11   Organ Weight:Body Weight Ratio	   28
  12   Organ WeightrBrain Weight Ratio 	   32
  13   Abnormal Changes Observed for Liver, Skin and Thyroid ....   36
  14   Grading of Deeree of Hyperplasia From Microscopic
         Appearance of the Thyroid Gland 	   37
  15   Thyroid Hyperplasia Observed in ETU Test Animals	   38
  16   Changes in Thyroid Morphology in Rats Which
         Ingested 625 ppm ETU for Ninety Days	   39
                                    vii

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                               INTRODUCTION
     The salts of ethylenebisdithiocarbamic acid (EBDC) have been exten-
sively used against a variety of pathogenic plant fungi.  A principal
impurity and degradation product of the EBDC fungicides is ethylene
thiourea (2-imidazolidinethione; ETU).   ETU has been identified as an
impurity in commercial EBDC formulations (1, 2).  It has been reported
to occur as a result of metabolic (3, 4, 5) and chemical (6, 7, 8)
alterations  of the commercial fungicides.   ETU has been identified on
a number of different crops which had been field-sprayed with a commer-
cial formulation of EBDC (9, 10).  Cooking of foods containing EBDC
residues also results in the formation of ETU (11, 12).

     Early studies evaluating the toxicity of the EBDC fungicides reported
a major effect to be thyroid hyperplasia and neoplasia (13, 14, 15).  The
toxicity  of ETU has also been studied and found to be essentially
similar to that of the parent fungicide, producing thyroid neoplasia in
rats (16, 17) and liver tumors in mice (14).  In these published reports,
recorded parameters consisted of body weights, organ weights, food con-
sumption, ^ I uptake, and terminal histopathology.  In the ETU subacute
toxicity study reported herein, thyroid hormone level measurements were
also made to determine at what dietary dose level ETU causes thyroid
hormone alterations and to establish a dietary concentration which does
not induce thyroid changes leading to hyperplasia.  In addition, it was
hoped that a relationship between altered thyroid hormone levels and
thyroid hyperplasia could be established.

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                                EXPERIMENTAL
                                 Materials

     Sprague  Dawley-derived cats* were purchased.front Charles River
Laboratories; (W^liningtBwi- Massachnsettsr)-..   E0J- (estimated? to?be-->95h
percent pure) was obtained front the: Aldrich Chemical Company  (Milwaukee,.
Wisconsin),, and prap§rlth±aurac±l  (FEU)  and antitroie- (3^-amino—l-,2,4—
triazole)' were- purchased? fram*- the1; Sigma* Chemical. .Company/  (St.. Louia^
Missouri)r.  1^1 f or i the- ehyraidt iodine* uptake: studies; was. obtained, from*
New England^Nueleat (Btkafcgm^Ma.R.<₯afthusefcfca).^ . CommprriaT,; test .,kfIrs, .fan
measuring- serum thyroxiner (fT~4^ an-d!. thg^cai-d- binding: g-lobuitnr (XB&^> were-:
purchased.front.Nueleat.Medical.  Laboratories» Inc.. (Dallas.,, Texas),^
rriiodothyronine- (T—1): Mts.werff. obtained, fixrar^DiagnastLCSLFrodiiets
Corporation^ (Ixss-Angeles.^,CaJifbJcmiaOfe.-  Initiai. serunn TSH, measurements^.
were perfG,Emed^wifehT>Mtsj pusEchased^faEomixBeckmait InstarametttSj.: Incr^.
(Fullerton>, eaMfd.rffia>.  Shortly afterr tMs? study began; the speeif±—
city of thei antiserufflT for'human? ISffi wasechangede andc. thfr-measttrfc o«f^ rafe .
TSH was no  longer accomplished..

     TSH kits from ICN'Medi-cai, Phaiina^arDia^iaffstics andt CaiLbioeheiih,
were purchased.and. evalua'ted as possible substitutes; for-measuring cat:
TSHv  Only  the: Galsbiachenb T'SBi RiEAL Mfc pxoxedL suitable,,, amdi. it. was= usede
at the end;  of this study.  TSH-was-not  measured during: most. o.£. the. study,..
                             ETU Purity-Analysig

     The ETIT used in, this study was: anaihy.zed by mass; sipecfeEoscopy and:
nuclear magnetic resonance .spectrometry- (NMR):».  Direct prober high
resolution, mass spectroscopy showed: a strong: parent ion. at,m/e 103.
This is shown in:. Figures: 1- and 2...   It wasi estimated: tharough  the-use.
both l^c and 2ft NMR. spectros;copy that ETU was 96..8. percent, pure..
                        Thyroid Euacticab and Coatrol.

     Thyroid hormone is. biosynthesized .in .the- thyroid: gland by; the
iodinatiom of tyrosine^  lodatyrosinea are coupled to_ fornL..T-3 and T-4.
These active hormones are~reieased into-rthe- bloodstream" and-are -dist-ri- --
buted throughout the-body where, they, regulate, metabolic., processes.
Hormone synthesis: and. release- is. eoatrfflled by a peptide which, originates
in the anfeer-iot lobe- ofc the. pituitary..,, the., thyroid stimulating, hormone

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(TSH).  Pituitary secretion of TSH is, in turn, regulated by two factors,
a hormone released by the hypothalamus called thyrotropin releasing
hormone (TRH) ,  and the concentration of free thyroid hormones (T-3 and
T-4) in the interstitial fluid which bathes both the pituitary and hypo-
thalamus.

     Increased serum thyroid hormone concentrations suppress secretion
of TSH while decreased hormone levels augment TSH release, providing for
a steady-state thyroid hormone concentration.  Recent studies indicate
that thyroid hormone blocks pituitary response to TRH.  As the serum
levels of T-3 and T-4 fall below a threshold concentration, the TSH-
secreting cells of the anterior pituitary become responsive to TRH and
secrete TSH which, in turn, raises serum thyroid hormone levels.  This
feedback control system is designed to provide a constant supply of
thyroid hormone to peripheral tissues, where it acts as a regulator of
cellular metabolism.

     Chemical substances that impair the synthesis or release of thyroid
hormone cause hyperplasia of the thyroid gland.  Continued exposure to
antithyroid chemicals results in the transformation of the hyperplastic
tissue to a malignant form.  It is now well documented that chemicals
altering thyroid function can initiate thyroid neoplasia (18, 19).
Chemicals which impair the synthesis or release of TSH from the pitui-
tary can severely affect the feedback control system which regulates
serum T-3 and T-4 levels.

     In summary, altered thyroid function can result from the action of
a chemical on iodine uptake and incorporation, synthesis or release of
thyroid hormones, synthesis or release of TSH, or inactivation of
receptors activated by either TSH (at the level of the thyroid) or by
thyroid hormone (at the pituitary level).  Using commercially available
test kits, the serum concentration of T-3, T-4, TBG and TSH can be
measured.   125 j uptake can be measured directly.  The Free Thyroxine
Index (FTI) was calculated and is reported for each group.  The FTI is
a measurement of free serum T-4.  While the T-4 level, measured by the
T-4 radioimmunoassay kit, is influenced by TBG concentrations, the FTI
is a measurement of the amount of T-4 free in the serum, independent
of the serum TBG concentration.  The formula used to calculate the FTI
is shown below:

                         Evaluation of Test Kits
     We have previously shown that certain chemically induced alterations
in thyroid function in rats could be measured using commercially avail-
able kits.  We reported serum T-4 and TBG (T-3 binding) levels and 125I
uptake by the thyroid (20) .  Prior to the initiation of this ETU subacute

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toxicity bioassay, we evaluated a new f-3 kit produced by  Diagnostic
Products Corporation (Los Angeles, California), and a TSH kit from
Beckman Instruments (Fullerton, California).  The T-3 diagnostic kit
quantitatively measures total serum T-3 by radioinununoassay.  One hundred
microliters of each serum sample (or standard) is mixed with an equal
volume of 125i labeled T-3.  Upon addition of the T-3 specific antibody,
the endogenous serum T-3 and the radioactive T-3 compete for binding
sites.  This reaction takes place in the presence of ANS (anion-8-anilino-
1-naphthaline sulfonic acid) which blocks the interaction of T-3 with
other serum proteins.  A second antibody, goat antirabbit gamma globulin,
is added to separate the free serum T-3 from the bound T-3.  This solution
is well mixed and incubated at 37°C for 15 minutes.  An ice cold solution
of 6 percent polyethylene glycol is then added to help precipitate the
antibody bound T-3.  Following centrifugation (3,500xg; 20 min.), the
supernatant containing the free T-3 is decanted and the antibody bound
T-3 precipitant is counted.  A five point standard curve is constructed
and serum T-3 levels are determined from this curve.

     The Beckman TSH diagnostic kit quantitatively measures total serum
TSH by radioimmunoassay.  The procedure is similar to that described
above for the T-3 test.  Serum from rats pretreated with amitrole was
evaluated for T-3 and TSH levels and this was compared to values obtained
from control serum.

     Unfortunately, after the ETU subacute tox'icity study began, Beckman
switched to a new antiserum with increased specificity for human TSH.
This kit would no longer measure rat TSH.  Therefore, TSH kits from other
commercial sources were purchased and evaluated as possible alternates.
The kit produced by Calbiochem contains a relatively nonspecific TSH
antiserum and will measure rat serum TSH levels.  Unfortunately, this
was not confirmed until near the end of the study and for this reason,
TSH values are available for only very few groups.
                                 Methods

     ETU, PTU, or amitrole was added to corn oil and then incorporated
into powdered rodent feed(l) using a twin shell tumble blender.  The
final corn-oil concentration in the blended f-eed was 1 percent.  The
test diets were prepared fresh weekly.  The dietary levels of ETU were
625, 125, 25, 5 and 1 ppm, PTU was 125 ppm and amitrole, 50 ppm.  These
dietary levels of PTU and amitrole were previously shown to produce
significant antithyroid effects (21, 22).  A control group received
powdered diet containing 1 percent corn oil.  Each of the seven dosage
groups contained 60 male and 60 female rats.  The control group contained
72 male and 72 female rats.  The animals were placed on study as out-
lined in Table 1.
    Ralston Purina Company, Richmond, Indiana.

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     At 30-day intervals, 10 rats of each sex from each test group were
sacrificed and serum T-3, T-4, TBG and TSH concentrations were measured.
These rats were also used for necropsy.  The remaining 10 rats of each
sex per group were used for thyroid ^->I uptake studies.  Each rat was
individually caged throughout the study.  Body weight and food consump-
tion was determined weekly.  Each rat was toe-punched at the start of
the study for permanent identification.

     The 10 tissues removed during necropsy are listed in Table 2.  Due
to excessive salivation and abnormal skin-hair coat development at the
625 ppm ETU level, skin and mandibular salivary glands were also removed
during necropsy from the rats which received the high diet.  The thyroid
gland and liver were examined microscopically in all study groups.  These
two tissues were fixed in formalin imbedded in paraffin, sectioned (6
microns) and stained with Hematoxylin-Eosin.  The remaining tissues were
stored in 10 percent buffered formalin.
                                 RESULTS

     The rats which received 625 ppm ETU showed clinical signs of
poisoning by the eighth day on study.  Excessive salivation, loss of hair
or failure to replace normal hair loss, rough and bristly hair coat, and
scaly skin texture was observed.  Ingestion of ETU was calculated and
found to range from 11.7 to 15.2 mg/day in female rats to from 13.0 to
17.3 mg/day in male rats.  Table P shows data on the weekly consumption
of ETU and other chemicals by all animals on study.  Fourteen of the 40
rats on the 625 ppm ETU 60-day study died between days 40 and 50 of the
study.  Only one rat from the 90-day group died.

     The rats receiving 125, 25, 5 and 1 ppm ETU, control diet, 50 ppm
amitrole or 125 ppm PTU showed no adverse clinical effects.

     Consistent with the observable clinical changes, several biochemical
changes were noted.  Rats ingesting ETU at 625 ppm showed a significant
decrease in iodine uptake.  The percent T-3 bound to TBG was lower than
normal at the 30- and 90-day evaluations.  Serum concentrations of T-3
and T-4 were also very substantially lower than the levels obtained for
the control rats.  The serum TSH concentration for the 625 ppm rats,
measured only at the time of the 30-day evaluation, was increased to more
than twice the serum TSH concentration observed for the control rats.
Significant differences in free T-4 were also found, as determined by
the FTI.

     The rate of iodine uptake, the percentage of T-3 bound to TBG, and
the serum T-3 concentration in the rats receiving 125 ppm ETU were not
significantly different from the control group.  However, a statistically
significant decrease in total serum T-4 concentration and in free serum

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T-4 measured at the 30-, 60- and 90-day evaluations was observed.  The
serum TSH concentration in the 125 ppm ETU rats at 30 days was approxi-
mately three times the concentration measured in the control rats.

     With one exception, there were ho significant measurable differences
in iodine uptake, T-3 binding to TBG or serum T-3, T-4 arid TSH levels in
the rats receiving ETU at either 25, 5 or 1 ppm for 30, 60 or 90 days.
The rats ingesting 25 ppm ETU for 60 days showed a statistically signifi-
cant increase in serum T-4 concentration.  This was not evident at 30 or
90 days.

     After 30 days of amitrole ingestion, very significantly decreased
serum T-3 and T-4 levels were observed.  The female rats from this group
also had a decreased percent of T-3 bound to TBG.  Both the male and
female rats had an increased serum TSH concentration.  In a previous
report (EBIS toxicity study), we showed that ingestion of 50 ppm amitrole
for four 'days resulted in a significant decrease in iodine uptake and
T-4 serum concentrations.  While the decreased T-4 levels are still found
after 30 days of amitrole ingestion, iodine uptake appears to have
returned to normal, probably as a result of biological compensation.  At
60 and 90 days, normal values for all of the thyroid function measure-
ments were noted.

     The rats which received PTU had decreased serum T-3 and T-4 con-
centrations at 30, 60, and 90 days.  The serum TBG level was affected
only in the 60- and 90-day test groups.  Since the PTU study was added
after the ETU study had started, TSH values were obtainable for the 60-
and 90-day PTU test groups.  Both groups had markedly elevated serum TSH
levels.

     Tables 3, 4 and 5 list the percent iodine uptake, TBG, T-3 and T-4
concentrations after 30, 60 and 90 days on study, respectively.  Also
included in these tables are the values for the amitrole and PTU posi-
tive controls.  TSH values are given for those test groups in which it
was measured.  The calculated FTI for each test group is shown in Table 6.

     Diet consumption for all groups is shown in Figure 3.  With the
exception of those rats receiving 625 ppm ETU and those ingesting 125 PTU,
there was no significant difference in feed consumption.  A 30 percent to
40 percent reduction in food consumption was observed in the rats on the
625 ppm ETU and 125 ppm PTU test diets, compared to control animal con-
sumption levels.  The average weekly feed consumption for each test group
with the standard deviation is listed in Table 7.

     The amount of ETU, PTU or amitrole consumed per week is shown in
Figure 4.  The average amount of each test chemical consumed along with
the standard deviation is listed in Table 8. .

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     Individual body weights were recorded weekly throughout the study.
 Table  9  summarizes  the body weight  data  (group mean ±  S.D.).
Figure 5 shows the weekly body weight gain for each dietary level.  Only
those rats which receive  625 ppm ETU or 125 ppm PTU test diets showed a
marked decrease in body weight gain.  No difference in body weight gain
was  observed for those rats which received ETU at 125, 25, 5 or 1 ppm,
or amitrole, as compared to the control group.

     At the conclusion of the study, gross necropsies were performed and
certain organs were removed and weighed (see Table 2).    Table 10 lists
the mean organ weights and standard deviation for each group.  There
were no significant differences in organ weights in those rats fed 25, 5,
1 and 0 ppm ETU.  However, thyroid hyperplasia was observed in all animals
which received 625 and 125 ppm ETU, 125 ppm PTU or 50 ppm amitrole in
their diet for 30, 60 or 90 days.

     The organ-weight-to-body-weight ratios were calculated (Table 11).
The spleen-to-body-weight ratio for the rats which ingested 625 ppm
ETU is significantly decreased while the ratios for the thyroid, brain,
kidneys, testicles and pituitary are significantly higher than the
corresponding control values.  Increased thyroid-to-body-weight ratios
were also found for the rats which received 125 ppm PTU, 50 ppm amitrole
or 125 ppm ETU (Table 11).

     Organ-weight-to-brain-weight ratios were also calculated (Table' 12).
no significant differences were found between rats receiving 125, 25,
5, 1 and 0 ppm ETU.   For those rats ingesting 625 ppm ETU, the ratio
of heart, spleen, kidney, adrenals  (female) and ovaries-to-brain-
weight was significantly decreased and the thyroid-to-brain-weight ratio
was significantly increased compared to control values.  The thyroid-to-
brain-weight ratios in the rats which received 125 ppm ETU were also
significantly increased in size at 30, 60 and 90 days.

     An increased thyroid-to-brain-weight ratio was the only significant
organ-to-brain-weight ratio change found in the 50 ppm amitrole test
animals.  The animals which received 125 ppm PTU for 30, 60 or 90 days
showed a very significant decrease in organ-to-brain-weight ratios for
heart, spleen, kidney, ovary, liver and adrenals.   These animals had a
very significant increase in thyroid-to-brain-weight ratios compared to
the control values.  The organ-weight-to-brain-weight ratios for the rats
which received amitrole and PTU are listed in Table 12.

     Gross examinations revealed the majority of the rats which received
ETU at 125 or 625 ppm, as well as those animals which received PTU or
amitrole for 30, 60 or 90 days had enlarged red thyroids.  Grossly
visible patterns of centrilobular congestion were present in a few random
rats fed amitrole or ETU.  A summary of gross changes is presented in
Table 13.

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     Microscopic examination  of tissues was restricted to thyroid and
liver.  The thyroids were graded for degree of hyperplasia using a slight
modification of the procedure of Astwood (23), and as defined in Table  14.
Varying degrees of thyroid microfollicular hyperplasia resulted at feeding
levels of ETU above 25 ppm (Table 15).  Relatively mild changes were noted
in animals fed 125 ppm ETU for 30 days as compared to the moderate changes
observed in the rats fed 625 ppm ETU for the same time period.  Marked
thyroid microfollicular hyperplasia and reduced colloid formation were
found  in animals fed amitrole or PTU for 30 days.  At 60 days, the rats
receiving the three highest levels of ETU (625, 125 and 50 ppm) and the
amitrole treatment group showed moderate thyroid hyperplasia as compared
to the 60-day PTU group which was more marked in its thyroid hyperplastic
response and reduced colloid production.

     Ninety days of dietary exposure to 625 ppm ETU also resulted in
microfollicular hyperplasia but not as severe as seen with PTU in the
same time period.  Numerous changes ranging from adenomatous hyperplasia
of follicles to what appear to be true solid adenomas were present in
the thyroids in the 625 ppm ETU group.  Table  16 categorizes the hyper-
plastic thyroids from the 625 ppm ETU test group into either solid
adenomas, cystic adenomas, adenomatous change or cystic-like follicles.
No adenomas were observed in the thyroid sections taken from the other
dietary groups.  No liver tumors werei observed in the rats at any dosage
level of ETU or in those animals which received PTU or amitrole.  The
majority of the livers examined microscopically had mild centrilobular
changes which can be described as follows.  Portal triad areas have a
smooth sheet of hepatocytes cut relatively across the nucleus and cyto-
plasm at about the same level giving well defined cell boundaries and
cytoplasmic proportions.  As one approaches the mid-zonal area and pro-
gresses into the centrilobular area of small central veins, the pattern
changes to less distinct cell borders of swollen cytoplasmic material
with fewer apparent nuclei.  The chord pattern is less distinct and.there
is a mild increase in small.polyploid nuclei.   The general appearance is
that of increased cytoplasm, fewer nuclei and less orderly hepatic chords.

     The data presented in this report show no significant measurable
adverse effects of dietary ETU at dosages of 5 ppm and below.  Signifi-
cant toxic responses, observed as altered thyroid function and changes in
thyroid morphology, resulted from the ingestion of ETU at 125 ppm or 625
ppm for 30, 60 or 90 days.  The observable effects at 25 ppm were noted
only at the 60-day interval and were not seen at 30 or 90 days.  Based
on these data, a dietary no-effect level in rats might be expected to be
25 ppm over the 90-day study.

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                                REFERENCES

 1.  Clarke, D. C., Baum, H., Stanley, E. L. and Hester, W. F., Anal.
     Chem. 23_, 1842-1844 (1951).

 2.  Bontoyan, W. R., Looker, L. B., Kaiser, T. E., Giang, P., and
     Olive, B. M., Survey of Ethylene Thiourea in Commercial Ethylene
     Bis Dithiocarbamate Formulations.  J. Assoc. Off. Anal. Chem.
     _55, 923-925 (1972).

 3.  Engst, R. and Schnaak, W., Investigations  of  the Metabolism of
     Fungicidal Ethylene Bisdithiocarbamates Maneb and Zineb.
     Z. Lebensm. Unters. Forsch. 134, 216-221  (1967).

 4.  Seidler, H., Haertig, M., Schnaak, W. and Engst, R., Metabolism
     of Certain Insecticides and Fungicides in the Rat. Nabrung 14,
     363, 373  (1970).

 5.  Truhaut, R., Fujita, M. Lich, N. P. and Chaigneau, M. Compt.
     Rend. Serie D. 276, 229-233 (1973).

 6.  Lopatecki, L. E. and Newton, W., The Decomposition of
     Dithiocarbamate Fungicides with Special Reference to the Volatile
     Products.  Can.J. Bot. 30, 131-138  (1952).

 7.  Fishbein, L. and Fawkes, J., Thin-layer Chromatography of
     Metallic Derivatives of Ethylene Bisdithiocarbamic Acid and Their
     Degradation Products.  J. Chromatogr. 19, 364-369 (1965).

 8.  Engst, R. and Schnaak, W., Residues of Dithiocarbamate Fungicides
     and the Metabolites on Plant Foods. Residue Rev. 52, 45-67 (1974).

 9.  Yip, G., Onley, J. H., and Howard, S. F. Residues of Maneb and
     Ethylene Thiourea on Field-Sprayed Lettuce and Kale.  J. Assoc.
     Off. Anal. Chem. J>4, 1373-1375  (1971).

10.  Newsome, W. H., Determination of Ethylene Thiourea Residues in
     Apples. J. Agric. Food Chem. 20, 967-969  (1972).

11.  Newsome, W. H. and Laver, G. W., Effects of Boiling on the
     Formation of Ethylene Thiourea in Zineb Treated Food.  Bull.
     Environ. Contam. Toxicol. 10, 151-154 (1973).

12.  Watts, R. R., Stroherr, R. W. and Onley, J. H., Effects of
     Cooking on Ethylene Bisdithiocarbamate Degradation to Ethylene
     Thiourea. Bull. Environ. Contam. Toxicol. 12,  224-228 (1974).

13.  Blackwell-Smith, R., Finnegan, J. K., Larson,  P. S., Sahyoun,
     P. F., Dreyfuss, M. L. and Haag, H. B., Toxicological Studies
     on Zinc and_Disodium Ethylene Bisdithiocarbamates.  J. Pharmacol
     Exp. Ther.  109, 159-166 (1953).

-------
REFERENCES (Continued)

14.  Innes, J.R.M., Valerio, M.,  Ulland, B. M., Pallotta, A. J.,
     Petrucelli, L., Fishbein, L., Hart, E. R., Falk, H. L.,
     Klein, M., and Peters, A. J.,  Bioassay of Pesticides and
     Industrial Chemicals for Tumorigenicity in Mice.  J. Natl.
     Cancer Inst. _4J2, 1101-1109  (1969).

15.  Andrianova, M. M. and Alekseev, I. V., Carcinogenic Properties
     of the Pesticides Sevin, Maneb, Ziram and Zineb.  Vopr. Pitan.
     2£, 71-74  (1970).

16.  Ulland,  B. M., Weisburger, T. H., Weisburger, E. K., Rice,
     J. M., and Cypher, R., Thyroid Cancer In Rats From Ethylene
     Thiourea.  J. Natl. Cancer Inst. ^£, 483-484  (1972).

17.  Graham,  S. L., Davis, K. J.,  Hansen, W. H., and Graham. C. H.,
     Effects of Prolonged Ethylene Thiourea Ingestion on the Thyroid
     of the Rat.  Fd. Cosraet. Toxicol. 13>, 493-499 (1975).

18.  Jukes, T. H. and Shaffer, C. B., Antithyroid Effects of
     Arainotriazole.  Science JL32, 296-297  (1960).

19.  Purves,  H, D., and Griebach, W. E., Thyroid Tumors in Rats
     Treated with Thiourea.  Br.  J. Exp. Path. 28, 46-53 (1947).

20.  Freudenthal, R. I.', Kerchner, G. A.', Persing, R. L.,
     Baumel,  I., and Baron, R. L., Subacute Toxicity of Ethylene-
     bisisothiocyanate Sulfide in the Laboratory Rat.  J. Toxicol.
     Environ. Health, In Press.

21.  Griessen, M. and Lemarchand-Beraud, T., Thyrotropin Secretion
     and Metabolism in Rats during Propylthiouracil Treatment,
     Endocinol. ^2_, 166-173 (1973).

22.  Seifter, J. and Ehrich, W. E., Goitrogenic Compounds:
     Pharmacological and Pathological Effects, 92, 303-314 (1948).

23.  Astwood, E. B., The Chemical Nature of Compounds Which Inhibit
     the Function of the Thyroid Gland.  J. Pharmacol. Exp. Ther.
     78, 79-82  (1943).
                                     10

-------
o
o
          OF spm.E era.
o
r
    0     10    20
                                                        fc MASS SPF'.n/r.MFi



                                                                   [n»at,    or
>r..
                                     iTiiii v. • eL V/ '-<;->ncfvjnf Go-



                                                    ___ -jV Jg




                                               ~*  Ar-p/voil
                                   Seen, .-...) f-
                                                              GC A t«r;
                                                                       v
         FIGURE 1.  MS ANALYSIS  OF THE ETU USED AS THE  TEST CHEMICAL
                                   11

-------
UlO
<£<*
U.O
OVj
  ift
  O _
      k
       "•"I""'
      CO   €0   70   0O
^0   100  110  ItO  130  110  ISO  ICO  170  180  190
              FIGURE 2.  MS OF THE ETU USED AS THE TEST  CHEMICAL
                                      12

-------
  260

  250

  240

  230

  220

  210

  200

  190

  180
O)
-170
o
o
160

150

140

130

120

110

100

 90

 80

 70

 60
            625 ppm   M
            —	T- T-
   A	A 625 ppm
                         Diet Consumption
                                 Amitrole
                               A	A  50 ppm   M
                               A	A  50 ppm   F
            125 ppm   M
-  D	D 125 ppm   F
         ••  25 ppm   M
                 I
                      I
                           I
                                      I
                                    j
           1234567
                                Weeks on Study
       FIGURE  3.   DIET  CONSUMPTION
                                                         10    11
                                                                12
                                    13

-------
                       Test Chemicgl Consumption
                625 ppm   M
       A	A 625 ppm   F
       Q     Q 125 ppm   M
       Q--r--p 125 ppm   F
       •...	-•  25 ppm   M
       9_	£  25 ppm   F
       #rrr—#   5 ppm   M
       #	#   5 ppm   F
       X.,   X   1 ppm   M
             -X   1 ppm   F
                  0 ppm   M
       Or--•-•<>   0 ppm   F
  Amitrole
A	-A  50 ppm   M
A	-4k  50 ppm   F
  PTU
         125 ppm   M
         125 ppm   F
45

40

35

30

25

20 |

15

10
                                        7     8
                             Weeks on Study
     FIGURE 4.   TEST CHEMICAL CONSUMPTION

                                 14
                                                        10   11    12

-------
   500
   450
   400
   350
»  30°
'5<
   250
   200
   150
   100
                                   Body Weight Gain
         O	O
  Amitrole
A	A  50 ppm  M
A	A  50 ppm  F
           PTU
                  125 ppm  M
                  125 ppm  F
            I
        I
1
I
I
1
                 1234567
                                    Weeks on Study
       FIGURE  5.   BODY  WEIGHT GAIN
                                           15
                                                        10
                                                11    12

-------
     TABLE 1.   ETU PROTOCOL DESIGN

ETU (ppm)
625
625
125
125
25
25
5
5
1
1
0
0
Amitrole
(ppm)
50
50
PTU
(ppm)
125
125
Sex
M
F
M
F
M
F
M
F
M
F
M
F


M
F


M
F
Number of
30 Days
20
20
20
20
20
20
20
20
20
20
24
24


20
20


20
20
Rats/Treatment Period
60 Days
20
20
20
20
20
20
20
20
20
20
24
24


20
20


20
20
90 Days
20
20
20
20
20
20
20
20
20
20
24
24


20
20


20
20

     TABLE 2.  TISSUES REMOVED AND WEIGHED
               DURING NECROPSY
Heart
Spleen
Kidney
Ovary (paired)
Testicle (paired)
*Liver
 Pituitary
*Thyroid (paired)
 Adrenal (paired)
 Brain
* Tissues processed for histopathologic evaluation.
                        16

-------
        TABLE 3.   THYROID  HORMONE  LEVELS  - AFTER  30 DAYS ON STUDY

ETU
(ppm)

625
625
125
125
25
25
5
5
1
1
0
0
Sex

M
F
M
F
M
F
M
F
M
F
M
F
125
I
(percent
uptake)

1.2 ±
2.1 ±
3.6 ±
4.0 ±
2.9 ±
3.2 ±
3.6 ±
3.8 ±
3.7 ±
3.0 ±
3.6 ±
3.5 ±
*
0.4*
1.4
0.8
1.7
0.6
1.3
0.6
1.0
0.7
0.5
0.9
0.9
TBG
(percent T-3
bound)

60.3
60.6
62.7
61.5
65.7
63.5
69.3
68.9
64.5
63.4
68.0
66.0
*
± 2.6*
± 1.8,
± 1.3,
± 1.3
± 2.3
± 2.0
± 6.3
± 1.3
± 1.2
± 1.3
± 5.6
± 5.2
T-3
(ng
percent)

57.3
58.4
71.1
104.4
67.1
86.3
79.0
88.1
82.1
90.9
76.0
83.2
A
± 3.7*
+ 9.9
±11.8*
±16.3
±15.9
±14.8
+ 8.1
±12.8
±13.0
±11.3
±11.8
±16.2
T-4
(ug
percent)

0.9 ±
1.1 ±
2.6 ±
2.1 ±
5.6 ±
3.8 ±
4.7 ±
2.9 ±
5.1 ±
3.5 ±
5.0 ±
3.8 ±
*
0.6*
1.0*
0.4
0.5*
1.1
0.8
0.4
0.9
1.0
1.0
1.7
1.4
TSH
(yiu
per ml)

1.4.3
14.6
23.3
18.3
7.3
5.1
6.7
4.9
6.4
4.5
6.7
6.0
*
± 0.9*
± 1-9*
± 5.9*
± 4.0
± 1.5
± 1.3
± 1.4
± 1.4
± 0.8
± 0.9
± 2.5
± 4.1
Amitrole
(ppra)
50
50
PTU
(ppm)
125
125

M 3.7 ± 2.1
F 5.0 ± 3.5


M 2.9 ± 1.1
F 3.3 ± 0.7

63.0 ± 1.5
61.4 ± 1.0*


67.8 ± 2.0
69.5 ± 1.6

*
49.1 + 19.5*
55. 2 ±


58. 9 ±
52. 0±
7.2

*
6.1*
8.0

0.
0.


0.
0.

6
3


9
7

± 0
± 0


± 0
± 0
*
•8*
.3

*
•2*
.1
*
9.3 ± 0.6*
8.9 ± 0.6


— -


Significantly different (p <0.05) from corresponding control.   Student's t
test was used to make comparison between the control and treated animals.
All data reported as the mean, ± S.D.
                                     17

-------
           TABLE 4.  THYROID HORMONE LEVELS - AFTER 60 DAYS ON STUDY
ETU
(ppm)
625
625
125
125
25
25
5
5
1
1
0
0
125
I
(percent
Sex
M
F
M
F
M
F
M
F
M
F
M
F
uptake)
1.9
2.4
3.6
3.3
3.2
3^7
3.5
4.0
2.7
3.2
4.3
3.5
+
+
+
+
+
+
+
+
+
±
+
+
1.0*
1.8*
1.4
1.0
0.7
1.3
0.8
0.8
0.6
0.7
0.9
0.8
TBG
(percent T-3
bound)
79.0
71.8
66.3
66,3
76.9
74.7
66.4
64.0
70.4
67.1
73.6
69.4
± 0.9
± 1.4
± 1.3
± 2.1
± 1.6
± 1.7
± 1.2
± 1.8
± 1.2
± 1.3
± 4.9
± 4.3
T-3
(ng
Percent)
56.9 ±
56.8 ±
79,8 ±
78.5 ±
86.4 ±
126.2 ±
85.4 ±
118,5 ±
80.3 ±
93.3 ±
77.3 ±
103,8 ±
10.3*
6.9*
28.1
28.6*
7.6
15.1
12.7
14.3
12.0
13.5
8.5
19.1
T-4
(yg
percent)
0.2
0.2
2.8
2.0
2.8
2.6
4.9
2.9
4.9
2.8
4.8
3.3
± 0.1*
± 0.1*
± 0.5*
± 0.5*
± 0.5*
± 0.5*
± 0.5
± 0.9
± 0.7
± 0.8
± 0.7
± 0.5
THS
(yiu
per ml)
_
-

-
-
-
-
-
-
-
5.8 ± 0.4**
6.4 ± 0.9**
Amitrol'e
(ppm)
50
50
PTU
(ppm)
125
125

M
F


M
F

4.0
4.8


3.9
5.4

± 0.7
± 0.9


± 1.6
± 1.7

74.
69,


61.
62.

8
8


7
2

± 1.6
± 1.3


± 2.6*
± 2.1*

83.4
111,6


46.1
50.9

± 8.9
± 10.7


± 3.9*
± 9.7*

5.9
3.5


1.2
0.8

±0.8
± 3.6


± 0.2*
±6.1*

—
-


9.8 ± 1.0*
10.8 ± 1.9*
*  Significantly different (p < 0.05) from corresponding control.
** TSH values to be used as control for PTU group.
                                         18

-------
          TABLE 5.   THYROID HORMONE LEVELS - AFTER 90 DAYS ON STUDY

ETU
(ppm)

625
625
125
125
25
25
5
5
1
1
0
0
Amitrole .
(ppm)
50
50
PTU
(ppm)
125
125
Sex

M
F
M
F
M
F
M
F
M
F
M
F


M
F


M
F
125
I
(percent
uptake)

2.5
3.7
2.8
3.9
3.3
3.4
3.7
4.2
3.5
3.2
3.8
4.1


5.8
4.8


4.7
5.6
*
± 0.8
±: 1.8
± 0.7
± 1.1
± 0.7
± 0.9
± 0.6
± 1.1
± 0.6
± 0.9
± 0.5
± 1.0


± 3.1
± 3.2


± 1.7
± 2.5
TBG
(percent
bound)

62
62
65
64
68
65
71
70
65
63
69
65


71
69


59
60

.7 ± 2.
.7 ± 0.
.3 ± 1.
.3 ± 1.
.9 ± 1.
.6 ± 2.
.4 ± 0.
.1 ± 2.
.8 ± 1.
.1 ± 1.
.3 ± 2.
.2 ± 2.


.4 ± 1.
.6 + 2.


.1 ± 1.
.4 ± 1.
T-3
*
o*
9
1
6
5
3
8
2
1
4
7
9


5
1

*
3*
7
T-3
(ng
percent)

27
35
86
105
79
108
76
105
68
116
72
106


67
94


73
69

.9 ±
.2 ±
.1 ±
.5 ±
.4 ±
.7 ±
.1 ±
.2 ±
.7 ±
.7 ±
.0 ±
.8 ±


.3 ±
.8 ±


.2 ±
.6 ±
*
13.3*
4.3
15.0
16.0
12.6
11.6
13.1
16.6
9.9
17.6
21.5
25.0


11.8
13.3


9-9*
9.4
T-4 TSH
(ug (uiu
percent) per ml)

1.1 ±
1.1 ±
2.3 ±
1.6 ±
3.8 ±
2.9 ±
5.0 ±
3.0 ±
4.0 ±
2.5 ±
4.5 ±
3.3 ±


3.6 ±
3.5 ±


0.6 ±
0.4 ±
*
0.6*
0.6*
0.6*
0.3
1.0
0.7
1.0
0.7
1.0
0-7 - . **
0.8 5.8 ± 0.4**
0.8 6.4 ± 0.9


1.3
1.1

* *
0.2* 9.4 ± 1.3*
0.2 10.7 ± 2.1

 Significantly different (p < 0.05) from corresponding control.


**
  TSH values to be used as control for PTU group.
                                            19

-------
TABLE 6.  FREE THYROXINE INDEX
-. . . - • •
ETU (ppm)
625
625
125.
125
25
25!
5
5
1
1
6
0
Ami;tr,ple
5Q-
50-,
PTU
(ppm)
125
125'
Sex
M
F
M
F
M
F
M
F
M-
F
M
F

U|
F,


M
F'
Days
30
0.5,2
0.67
1.6.2
1.27
3.69;
2, 42*
3.24,
2'. OP.
3 ."28.
2.22
3 . 3.7
2.52.

Q>3&
0.15,


0.64.
0.49-
on Study
6Q
Q.15
0.12
1,86
1.35
2'..12
1.9;i
3:.2-7-
1 ..88;
3.,49
1.86>
3'.52'
2...31:

4, 37
2 .42


0, 74,
0,48;
:90
0.68
0.70
1.4,8
1,0.5
2. .64
1 . 9,0.
3.. 5.3:
2.12
2.64.
1.52:
3j. 12
2. .14

2.53
2.. 41


0.33',
0,23
              20

-------
TABLE 7.  WEEKLY DIET CONSUMPTION

ETU
(ppm)
625
625
125
125
25
25
5
5
1
1
0
0
Amitrole
(ppm)
50
50
PTU
(ppm)
125
125
Sex
M
F
M
F
M
F
M
F
M
F
M
F


M
F


M
F
Week
No. 1
91.2 + 14.1
85.0 + 14.9
120.3 + 27.0
105.8 + 33.5
114.7 + 27.0
95.0 + 20.8
136.2 + 18.9
121.6 -1- 24.8
137.0 + 18.0
112.3 + 13.6
145.5 + 35.2
123.3 + 43.3


138.4 + 16.4
114.0 + 19.2


150.6 + 17.7
120.7 + 13.8
Week
No. 2
114.6 + 29.5
103.8 + 30.7
141.9 + 19.5
107.7 + 18,7
153.6 + 17.0
119.8 + 22.9
155.4 + 20.5
115.4 + 23.2
143.5 + 26.0
116.5 + 23.2
151.1 + 20.4
119.1 + 22.8


146.1 + 17.6
118.8 + 20.8


136.7 + 17.5
108.6 +16.6
Week
No. 3
102.0 + 14.8
83.9 + 24.3
152.7 + 20.6
115.8 + 22.9
165.3 + 23.6
129.8 + 37.9
167.9 + 26.8
123.1 + 19.9
171.1 + 28.1
123.1 + 27.9
169.9 + 21.5
131.7 + 43.2


162.3 + 19.8
127.3 + 31.7


113.6 + 23.4
95.5 + 23.7
Week
No. 4
100.7 + 16.3
84.0 + 17.2
150.6 + 27.3
126.8 + 42.2
166.6 + 18.7
127.1 + 27.6
171.2 + 36.1
129.3 + 38.7
173.9 + 41.5
121.2 + 29.3
168.0 + 20.3
131.6 + 39.1


165.6 + 19.9
133.3 + 32.9


100.1 + 17.2
74.4 + 13.3
Week
No. 5
98.5 + 17.2
82.2 + 24.1
171.3 + 35.2
136.8 + 58.3
169.2 + 13.6
116.6 + 17.5
169.0 + 21.1
128.7 + 29.3
168.8 + 13.4
120.9 + 23.3
176.7 -1- 35.0
140.9 + 39.7


175.8 + 26.0
130.2 + 26.9


104.1 + 14.5
86.7 + 27.6
Week
No. 6
101.0 + 20.8
85.2 + 23.9
159.6 + 22.1
124.5 + 31.4
172.1 + 24.9
121.1 + 25.1
174.3 + 19.8
128.8 + 20.7
179.0 + 27.3
140.3 + 43.4
182.9 + 47.7
146.3 + 59.2


169.9 + 30.9
127.1 + 26.8


96.8 + 18.7
89.1 + 21.2

-------
                                               TABLE 7.   (Continued)
ro
ts)
ETU
(ppm)
625
625
125
125
25
25
5
5
1
1
0
0
Amitrole
(ppm)
50
50
PTU
(ppm)
125
125
Sex
M
F
M
F
M
F
M
F
M
F
M
F


M
F


M
F
Week
No. 7
107.8 + 17.9
92.6 + 24.8
165.0 + 13.7
114.7 + 11.9
188.7 + 43.9
138.3 + 32.9
173.6 + 27.7
125.2 + 26.9
181.9 + 45.9
144.6 + 53.2
191.6 + 48.7
146.8 + 48.7


177.6 + 40.8
138.0 + 51.6


102.7 + 19.0
95.5 ± 19.5
Week
No. 8
117.9 +15.3
105.2 + 20.8
178.2 + 43.2
120.5 +21.2
183.5 + 23.1
139.0 + 43.3
181.0 + 37.4
133.9 + 51.9
182.9 + 33.2
131.9 + 34.3
181.8 + 48.7
146.6 + 55.8


182.1 + 38.0
132.9 +48.4


96.4 + 18.9
91.3 + 28.1
Week
No. 9
117/8 + 20.7
106.1 +27.1
167.6 +38.4
163.1 + 83.1
195.3 + 34.4
143.3 + 36.9
173.2 + 31.2
130,7 + 25.4
182.7 + 27.9
146.0 + 46.7
177.8 + 30.5
138.1 + 36.2


185.9 + 39.9
129.2 + 40.8


91.0 + 19.4
96.5 + 28.3
Week
No. 10
121.0 + 20.1
101.8 + 20.8
176.7 +29.5
146.6 + 54.8
176.8 + 16.3
133.0 + 26.1
158.6 + 35.3
127.6 + 47.0
207.8 + 56.2
145.8 + 49.1
186.5 + 40.4
153.2 + 66.6


229.1 + 75.6
174.6 + 79.8


101.9 + 16.5
89.1 + 20.8
Week
No. 11
114.8 + 17.2
93.8 + 13.5
181.2 + 46.8
137.1 + 50.9
198.2 + 51.7
151.4 + 63.1
172.9 + 38.3
129.7 +24.3
207.9 + 54.2
148.1 +61.5
183.4 + 45.0
149.0 + 66.8


165.8 + 22.0
118.4 + 25.7


91.5 +18.1
68.5 + 9.9
Week
No. 12
91.6 + 18.7
89.8 + 30.2
164.3 + 27.9
129.6 + 56.9
200.7 + 59.8
170.0 + 69.4
154.2 + 18.0
111.1 + 16.5
168.8 + 18.0
109.6 + 14.9
177.8 + 46.2
131.7 +50.9


192.5 + 32.1
138.1 + 59.0


92.0 + 14.1
88.2 + 15.4

        Mean,  + S.D.;  presented in grams,

-------
                                       TABLE  8. WEEKLY TEST CHEMICAL INGESTION*
ro
OJ

ETU
(ppm)
625
625
125
125
25
25
5
5
1
1
0
0
Amitrole
(ppm)
50
50
PTU
(ppm)
125
125
Sex
M
F
M
F
M
F
M
F
M
F
M
F


M
F


M
F
Week
No. 1
57.0
53.1
15.0
13.2
2.9
2.4
0.7
0.6
0.1
0.1
0
0


6.9
5.7


18.8
15.1
Week
No. 2
71.6
64.9
17.7
13.5
3.8
3.0
0.8
0.6
0.1
0.1
0
0


7.3
5.9


17.1
13.6
Week
No. 3
63.7
52.4
19.1
14.5
4.1
3.3
0.8
0.6
0.2
0.1
0
0


8.1
6.4


14.2
11.9
Week
No. 4
62.9
52.5
18.8
15.9
4.2
3.2
0.9
0.7
0.2
0.1
0
0


8.3
6.7


12.5
9.3
Week
No. 5
61.6
51.4
21.4
17.1
4.2
2.9
0.9
0.6
0.2
0.1
0
0


8.8
6.5


13.0
10.8
Week
No. 6
63.1
53.3
19.9
15.6
4.3
3.0
0.9
0.6
0.2
0.1
0
0


8.5
6.4


12.1
11.1
Week
No. 7
67.4
57.9
20.6
14.3
4.7
3.5
0.9
0.6
0.2
0.1
0
0


8.9
6.9


12.8
11.9
Week
No. 8
73.7
65.7
22.3
15.1
4.6
3.5
0.9
0.7
0.2
0.1
0
0


9.1
6.7


12.1
11.4
Week
No. 9
73.6
66.3
21.0
20.4
4.9
3.6
0.9
0.7
0.2
0.2
0
0


9.3
6.5


11.4
12.1
Week
No. 10
75.6
63.6
22.1
18.3
4.4
3.3
0.8
0.6
0.2
0.2
0
0


11.5
8.7


12.7
11.1
Week
No. 11
71.7
58.6
22.7
17.1
5.0
3.8
0.9
0.7
0.2
0.2
0
0


8.3
5.9


11.4
8.6
Week
No. 12
57.3
56.1
20.5
16.2
5.0
4.3
0.8
0.6
0.2
0.1
0
0


9.6
6.9


11.5
11.0

       Mean,  + S.  D.;  presented in grams.

-------
                                        TABLE 9.  WEEKLY BODY WEIGHT GAIN
to
Weeks on Study
ETU, ppm
625
625
125
125
25
25
5
5
1
1
0
0
Amitrole
50
50
PTU
125
125
Sex
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
0
129.4 ± 14.2
117.5 ± 12.0
132.1 ± 17.5
116.61 14.6
145. 0± 11.3
127.2 ± 12.1
140.5 ± 13.9
124. 0± 12.4
131. 0± 16.6
115.2 ± 13.7
141. 0± 27.6
119. 0± 21.1
139.1± 18.7
120.21 13.7
168.5+ 39.8;
136.41 26.6
1
141.4 H5.0
127.2 ±12.6
180.3 ±22.7
146.3 ±13.1
188.6 ±18.1
152.1 ±13.3
195.3 1 14.4
155.1 ±11.4
191.5 ± 22.1
151.6 1 13.9
193.8 1 29.4
151.61 18.0
199.51 20.8
155.51 16.2
224.31 35.2
167.81 22.5
2
157.7 117.8
135.5 112.6
226.7 ±24.4
168.2 ±15.1
242.1 115.2
176.5 +14.8
248.9 ± 21.5
179.4 ±23.9
234.01 29.9
173.4 1 14.7
242.81 30.1
174.91 22.2
245.31 25.7
176.71 19.0
241.91 37.0
174.11 24.3
3
172.ll 24.3
143.61 15.8
274.01 27.0
188.91 16.0
295.01 20.0
199.71 16.8
299.81 22.9
201.61 19.6
292.71 32.7
193.81 17.3
290.5 +36.9
194.5 ±21.5
291.5 + 27.5
195.1 ±23.7
245.3 + 34.9
176.3 ± 25.8
4
187.2 ± 25.3
148.6 ±17.5
306.6 ±37.3
204.1 ± 20.8
335.9 ±21.9
217.4 ±.18.4
340.1 ± 28.8
218.2 ± 21.5
332.3 1 35.2
213.31 20.2
331. 9± 35.8
211.01 20.1
328.61 27.2
216.51 21.6
247. 4± 32.1
178. 1± 24.2
5
195.1 ±21.1
152.6 116.2
341.3 ±38.2
217.3 ±25.8
376.9 ±22.9
228.8 ± 20.6
373.9 ± 30.4
238.5 ± 22.8
377. 5± 37.6
227.4± 17.8
366.4+ 35.3
228.9+ 21.6
361.6+ 30.5
232.6+ 24.8
261. 7± 30.2
183. 5±. 26. 9
6
189.6 ±26.5
145.7 ±21.5
365.7 ±38.0
230.7 ±23.0
401.3 ± 21.3
241.0 ± 23.8
398.5 ± 36.3
254.1 ± 22.0
408. 6± 40.9
238. 1± 18.4
394.5+ 33.6
242.3±;23.7
378. 3± 37.5'
243.4+ 23.9
265.0+ 29.3
185.0+ 26.7

-------
                                                TABLE  9.   (Continued)
ETU, ppm
625
625
125
125
25
25
5
5
1
1
0
0
Amitrole
50
50
PTU
125
125
Sex
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F

7
195.7 ±36.0
148.8 ± 25.0
385. 3± 40.5
242. 1± 23.4
420. 1± 29.0
250. 9± 22.9
425.1+ 39.4
263. 3± 24.1
428. 6 ± 44.4
249. 1± 19.6
417. 9± 36.0
251. 1± 22.0
407. 4± 37.0
249. 8± 22.7
265. 5± 30.4
184. 9 ± 27.5

8
210.7 ± 34.6
161.9 ± 21.6
413.4 ±32.3
249. 9± 26.6
443.0+ 26.0
254.3 + 25.6
447. 8± 42.2
269. 6± 25.4
446.7+ 49.7
256.5+ 19.1
432.9+ 39.7
263. 3± 30.6
428. 8± 44.2
258.2+ 25.8
265.7+ 31.0
183.71 27.8
Weeks on
9
234.8 ±29.8
170.3 +25.7
431.4 ±36.7
250.8 ±22.0
451.9 +29.2
273.6 +25.0
457.7 +38.4
274.9 +26.5
442.0 ±49.0
260,0 ±20.8
448.9 ±35.7
266.6 +30.5
437.5 ±44.4
254.9 ±32.8
259.0 ±33.0
171.8 ±28,0
Study
10
246.6 ±34.8
180.7 ±23.1
444.6 +40.2
254.5 +25.5
466.4 ±26.1
275.2 ±23.1
468.7 ±35.4
286.7 +25.8
462.0 ±49.4
267.5 ±20.1
461.1 ±38.8
275.2 ±26.6
456.4 ±40.1
263.8 ±31.2
259,3 + 35,5'
171,1 +28.9

11
25.1.2 ±38.2
181.3 ±23.9
462.5 ±40.9
265.7 ±24.4
483.5 ±33.1
283.3 +23.3
478.8 ±38.5
290.2 ±23.4
476.0 ±52.9
269.8 ±24.0
478.4 ±33.7
280.5 +28.0
471.1 ±40.0
269.0 ±32.2
265,3 + 35,7
172.4 +27,6

12
239.4 ±40.3
171.5 ± 28.3
472.1 ±43.3
268.1 ±23.4
497.5 ±33.8
288.8 ±26.9
491.9 ± 39.2
295.2 ±26.8
492.3 ± 55.3
276.3 ± 22.2
492. 1± 35.2
285.9 ± 30.7
487.7 ± 40.4
275.5 ± 32.6
265, 6 ± 37.2
174,5 ± 29.0
* Mean,  + S.D.; presented in grams.

-------
TABLE  10.  OKCAN WKICHTS DB'iAINEU AT NKCROFSY
F.TU
(ppm)
625
625
125
125
25
25
5
5
1
1
0
0
625
625
125
125
25
"' 25
5
5

1
0
0
625
62'.

125
25
25

r(
1
1
0
0
Days on
Study
90
90
90
90
90
90
90
90
90
90
90
90
60
60
60
60
60
60
60
60
60
60
60
60
10
10
1(1
to
11)
10
10
10
30
10
10
30
Heart
S>>x (K)
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
j,.
M
F
M
F
M
F
M
F
M
P
0.77
0.58
1.37
(1.88
1.41
0.97
1 .41
0.40
1 . 39
0.84
1.41
0.91
0.77
0.57
1.37
0.95
1.38
0.8.S
1.43
0.92
1.29
0.81
1.15
0.90
0.61
0. •>:>
1 .(IS
(1.8 t
1.1'.
O.K.'
1 . I 1
0.77
1.16
0.74
1.2)
0.86
' 0 . 1 ft
1 0. 1 1
' 0.1)
1 0.07
.1 1) . 1 4
1 I). 11
' 0.22
' 0.09
i 0. 15
! 0.06
. 0.19
'. 0.09
+ 0.31
±0.06
± 0.23
± 0.33
i 0.09
± 0.1 1
± 0.18
i 0.13
< 0.16
± 0. 10
1 0.21
' 0.09
i 0.08
i 0.118
i 0.09
i (1.08
i (1.12
. 0.07
i 0.09
. 0. 10
. 0. 12
1 0.05
i 0.26
' 0. 17
Si
.1.28
0.24
0.81
0.51
0.84
0.5)
(1.88
0.54
0.80
0.52
0.90
0.55
0.31
0.21
0.76
0.54
0.80
0.51
(1.84
0.54
0.72
0.55
0.74
0.56
ll.:)4
(1. 28
II.7/.
11.51
11.79
(1.54
11.79
II. 52
0.72
0.51
0.77
0.51
1 fun
(s)
i 0.11
' 0.07
' 0.11
' 0.12
> 0.12
' 0.09
1 I). 1 1
' 0.09
! 0.15
i 0.09
L 0.10
1 0.11
± 0.19
± 0.03
± 0.13
± 0.08
±0.10
±0.04
i 0.10
±0.07
± 0.09
±0.07
i 0.12
± 0.10
1 1.07
. 1.06
. I. 1
• 1.64
.1.11
i 1. Ill
i 1.08
. 1.06
t II. 11
t 11.08
±0.10
±0.06
Rt . Kidney
0.85
0.65
1.69
0.91
. 59
.00
.49
.00
.55
0.94
1.62
1.01
0.86
0.67
1.46
0.94
1.64
0.99
1.65
0.96
1.55
0.99
1 .64
1 .05
11.10
0.64
1 . 26
0.40
1 .29
0.92
1 .13
0.81
1.26
0.81
1.39
0.85
' 0. 1 1
' O.Od
' 0.29
' 0.10
1 0. 17
' II. Ill
' 0.19
' 0.09
' 0.22
1 0.08
! 0.20
'. 0. 1.1
i 0.19
1 0.03
i 0.08
i 0.06
1 0.21
i 0.14
'. 0.21
.' 0.07
' 0.19
1 0. 17
' 0.25
' 0.26
' 11. 1 2
' (1.09
1 0.09
1 0.09
' 0.119
' 0.119
1 0.15
' 0.09
1 0.08
' 0.05
' 0.18
1 0.07
l.oft Kidney
II. SI i
0.1.1 '
1.6'. .
0.92 i
1.61 .
0.98 i
1.44 i
0.97 i
1 .55 i
11.94 i
1 . 60 .
0.97 i
0.88 ±
0.65 ±
1.45 ±
0.91 ±
1.63 i
0.99 +
1.59 i
0.91 i
1.57 ...
0.94 i
1.62 ..
0.94 i
11.79 i
11.65 '
1.2'. i
11.86 '
1 .28 i
0.91 '
1.1'. '
0.82 i
1 .24 i
0.81 .'
1 . 18 i
0.85 .'
0.14
II.O5
0.11
0.08
0. 17
(1. 1 1
1). 18
II. 10
0.22
0.09
0.18
0.10
0.21
0.01
0.11
0.08
0.21
0.12
0.22
0.09
0.18
0.09
0.22
0.15
0. 12
0. Ill
O.ll'l
0. 10
0.07
(1.09
0. 11
0.08
0.08
0.06
0. 19
0.09
Ovary
(pal rod; K)

(1.08

11.09

I). 1(1

0. 1 1

0. 10

0.10

0.08

0.13

0.10

0.10

0.10

0. 12

0.09

0. 12

0. 12

0. 1 1

0.12

0.12

1 0.02

' 0.-02

' 0. 14

1 0.01

! 0.02

± 0.02

± 0.02

' 0.01

'. 0.02

1 0.02

1 0.02

' 0.01

1 0.01

' 0.02

1 0.02

1 0.01

1 0.02

•': 0.02
Tos
(pal
4.37

5.17

5.11

5 . 29

5.11

5.67

3.17

4.59

5.35

5.04

4.99

4.90

1.8 )

4 . 08

4.21

4.13

3 .16

4 . 23

tide
ri-d; K)
i 0

' 0

i 0

< 0

.' 0

± 0

± 1

+ 0

60

V.

71

55

10

55

14

51

' 0.53

± 0

± 0

. 0

1 0

i ()

1 ()


60

55

72

16

''4

4 5

i [),27

. g

f o

41

47

7
(,
19
9
17
10
16
10
17
10
17
10
8
7
17
10
20
10
17
111
16
9
J7
10
8
7
1 .5

14
4
16
8
14
Q
15
9
Liver
(R)
5d
51
06
78
62
92
62
72
98
57
33
21
00
57
42
12
82
32
86
53
69
49
21
11
16
92
72
44
88
7f,
08
83
18
64
24
92
i 1
. 0
1 -J
< I)
1 1
1 1
i 2.
• 1
i 1
< 1
. 1
. 1
i 1
± o
± 1
± 1
± 3
t 1
i 1
. .1
i 2
54
8 t
40
99
91
01
14
67
04
28
92
22
13
66
37
31
22
55
09
03
10
i 0.96
i 2
' 2
1 0

i .
1 0
1 1
1 0
1 2

1 1
1 0
i 1

58
24
88
2'.
15
95
I |
72
4 1
I 3
31
97
21
29
Pituit
(mtf
10
11
10
1 2
10
10
25
80
80
38
ary
)
! 1
• 4
i 2
1 2
'- 2
1470 ! 2
1 1
14
9
.11
11
14
10
9
13
12
10
8
8
11
9
12
10
16
12
7
10
10
8
8
8
a
7
7
9
11
(10
20
80
90
42
83
20
11
70
10
40
80
60
20
20
40
08
75
10
10
40
60
70
40
90
20
70
70
58
50
' 3
1 3
! 4
* 4
i 3
i 6
i 3
i 2
± 2
± 2
± 1
± 3
t 2
i 2
• 1
1 3
' 3
'- 6
1 •)
1 2

, 2

, 3

, 2
, 1
+ 2
7.5
^ 3
Thvrold
('««)
6 1
40
10
94
33
75
27
68
37
15
09
44
11
12
79
18
58
08
07
30
99
81
61
82
64

78
72
77
86
45
1 5
89
58
91
06
48
45
17
J. I
24
20
2'i
18
27
2 '
.in
< '
57
38
37
27
14
23
20
18
27
21
29
21
29
28
11
23
26
2.1
20
16
19
14
24
16
JO
!!S
.11
71
51
21
71
91
51
II
42
00
60
11
10
30
50
00
40
to
00
80
17
25
10
80
20
80
00
90
to
50
40
80
17
00
' 24
' 12
' 8
• 1
' 9
' 6
' 4
6
' 4
' 4
' 8
' 4
' 19
' 18
± 10
± 7
± 7
'- 5
i 4
i 4
± 6
i 6
i 13
i 9
i 8
'. 6
1 12
'. 7
i 6
.': 4
.' 4
1 3
-' 3
1 2
± 8
i 3
34
98
21
53
88
05
97
06
77
47
64
45
09
40
93
01
37
79
40
12
38
14
72
66
04
84
70
24
62
38
04
66
24
90
13
52
Adrona 1
(paired; m>>)
40.
43.
48.
52
51
59
48
61
49
64
51
66
41
41
55
63
48
59
47
63
50
57
52
65
38
37
45
58
48
61
49
55
45
61
45
57
70
13
20
10
30
60
90
60
80
40
13
08
40
00
90
00
50
40
60
40
00
10
25
00
00
20
40
20
40
30
70
50
30
30
50
75
'6.15
1 8.64
1 10,02
' 7.80
' 8.18
'- 8.1.9
i- 9.40
' 8.51
1 7. 11
' 10.27
i 6.10
±11.85
± 11.02
± 7.48
1 9.64
± 10.74
± 6.17
± 1.5.28
± 9.08
± 8.06
± 8.99
1 14.91
± 9.96
± 14.57
± 7.60
± 8.01.
'. 6.82
1 9 . 00
-' 11.61
J 6.62
± 8.72
i 5.87
i 7 . 66
± 11.56
i 6.57
± 6.21
Brain
(g)
1.80 ±
1.66 1
2.05 ±
1.81 ±
2.06 i
1.90 ±
2.05 ±
1.89 ±
2.04 ±
1.80 ±
2.08 ±
1.81 '
1.76
1.65
2.00
1.85
2.04
1.88
1.95
1.88
1.99
1.84
2.06
1.'90
.1.71
1.68
1.88
1.78
1.88
1.79
1.97
1.76
1.92
1.79
1.91
1.81
0.08
0.08
0.14
0.09
0.06
0.08
0.12
0.81
0.13
0.18
0.09
0.10
t 0.07
± 0.10
± 0.13
± 0.11
± 0.07
± 0.07
± 0.15
± 0.10
± 0.14
± 0.13
± 0.13
± 0.07
± 0.11
± 0.09
± 0.07
± 0.07
± 0.09
± 0.07
i 0.12
± 0.07
± 0.11
± 0.06
± 0.08
± 0.08

-------
TABLE 10.  (Continued)
Ami t role
(ppm)
50
50
50
50
50
50
PTU
(ppm)
125
125
125
125
125
125
Control
(ppm)
0
0
0
0
0
0
Days on
Study
90
90
60
60
30
30

90
90
60
60
30
30

90
90
60
60
30
30
Sex
M
F
M
F
M
F

M
F
M
F
M
F

M
F
M
F
M
F
Heart
(K)
1.29
0.89
1.27
0.92
0.89
0.72

0.71
0.54
0.73
0.62
0.60
0.52

1.43
0.93
1.35
0.90
1.23
0.86
* 0.15
k- 0.09
1 0.21
1 0.12
1 0.07
f 0.11

1 0.11
k 0.07
i 0-09
± 0.12
± C OS
± 0.07

t 0.19
t 0.09
1 0.21
k 0.09
1 0.2(.
1 0.17
Spleen
0.80
0.61
0.75
0.57
0.55
0.54

0.25
0.20
0.35
0.28
0.31
0.25

0.90
0.55
0.74
0.56
0.77
0.53
+ 0.11
1 0.09
^ 0.08
1 0.06
1 0.06
1 0.11

1 0.05
> 0.03
k 0.07
* 0.05
± 0.05
± 0.04

<• o.io
<• 0.11
< 0.12
1 0.10
1 0.12
t 0.06
Rt. Kidney
1.49
1.01
1.58
1.01
1.09
0.80

0.77
0.60
0.94
0.69
0.78
0.61

1.62
1.01
1.64
1 .05
1.39
0.88
f 0.20
! 0. II
1 0.21
1 0.08
t 0.08
1 0.06

1 0.06
1 0.10
t 0.10
t 0.06
± 0.10
t 0.07

J- 0.20
1 0.11
t 0.25
1 0.26
1 0.18
1 0.07
Left Kidney Ovary Testicle
(g) (paired; g) (paired; g)
1 .51
1.00
1.56
0.99
1.11
0.80

0.76
0.59
0.94
0.70
0.77
0.59

1.62
0.97
1.62
0.94
1.38
0.85
k- 0.
1 0.
1 0.
1 0.
1 0.
1 0.

1 0.
k 0.
t 0.
± 0.
± 0.
* 0.

0.
0.
0.
0.
0.
0.
19 -- 5.15 ± 0.45
11 0. 10 1 0.01
20 — 5.02 <" 0.54
11 0.10 1 0.02
08 -- 4.10 * 0.43
06 0.13 k 0.03

07 -- 4.47 l 0.34
1 1 0.07 ± 0.01
11 -- 4.84 -k- 0.62
09 0.09 t 0.02
09 -- 3.86 ± 0.18
05 0.09 ± 0.02

18 -- 5.67 + 0.55
10 0.10 * 0.02
22 — 4.90 ± 0.72
15 0.12 1 0.03
19 -- 4.23 > 0.47
09 0.12 t 0.02
17.
10.
17.
10.
13.
10.

8.
6.
9.
7.
8.
6.

Liver
25 '•'
48 1
33 k
53 t
73 »
70 '

39 ±
53 t
54 k
48 ±
01 ±
92 t

1 7. 33 k-
10.
17.
10.
15.
9.
21 k
23 1
33 *
24 '
92 '
2.70
0.76
2.79
1.27
2.30
1.85

0.89
1.19
1.31
0.97
1.40
0.84

1.92
1.22
2.58
2.24
1.21
l.l!9
Pituitarv
(mg)
12.44 t
17.30 k
10.40 k
14.60 k
7.80 i
7.60 *

8.11 i
9.90 ±
7.90 ±
8.90 ±
7.30 ±
7.20 ±

11.42 ±
14.83 k
10.08 *
16.75 k-
9 . 58 >
11.50 t
2.40
5.00
2.67
4.99
2.90
1.78

3.02
2.47
2.02
3.25
3.86
2.70

3.09
6.44
3.63
6.62
5.93
3.06
Thyroid
(paired; mg)
64
41
36
24
71
53

113
86
101
90
55.
62,

30,
25.
29.
23.
24,
16.
.80 k
.60 1
.30 k
.30 1
.90 1
.80 1

.40 ±
.10 ±
.80 ±
.20 =k
.60 ±
.20 ±

,42 t
,00 t
.17 I
.25 k
.17 k
,00 k
15.34
9.77
11.27
6.18
12.31
9.20

44.90
31.56
33.68
26.64
12.77
17.94

8.64
4.45
13.72
9.66
8.13
3.52
Adrenal
(paired; mg)
48.
61.
44.
67.
35.
52.

28.
26.
28.
34.
23.
27.

51.
66.
52.
65.
45.
57.
10 t
10 t
80 *
20 I
60 k
80 I

67 ±
60 ±
70 ±
20 ±
50 ±
70 ±

33 t
08 +
20 t
00 (-
50 k
75 +
8.35
7.20
7.22
10.59
8.02
7.90

6.67
3.63
5.96
5.41
2.07
5.72

6.30
11.85
9.96
14.57
6.57
6.21
Brain
(g)
2.01 +
1.90 k
2.03 *
1.87 t
1.89 t
1.72 t

1.85 ±
1.77 ±
1.92 t
1.80 ±
1.80 ±
1.71 ±

2.08 ±
1.83 ±
2.06 ±
1.90 *
1.91 ±
1.81 t
0.08
0.05
0.15
0.05
0.07
0.04

0.13
0.11
0.13
0.11
0.08
0.05

0.09
0.10
0.13
0.07
0.08
0.08

-------
TABLE 11.  ORGAN WEIGHT:BODY WEIGHT RATIO
ETU
(ppm)
625
625
125
125
25
25
5
5
1

1

0

0
625
625

125

125

25
25

5

5

1
1
0
0
625
625

125
125

25

25

5

5
1

1

0
0
Days
on
Study
90
90
90
90
90
90
90
90
90

90

90

90
60
60

60

60

60
60

60

60

60
60
60
60
30
30

30
30

30

30

30

00
30

30

30
30
Sex
M
F
M
F
M
F
M
F
M

F

M

F
M
F

M

F

M
F

M

F .

M
F
M
F
M
F

M
F

M

F

M

F
M

F

M
F
Heart
(g)
3.63 x 10"3
3.A4 x 10~3
2.82 x 10~3
3.25 x 10~3
2.78 x 10~3
3.29 x 10~3
2.85 x 10~3
3.13 x 10~3
2.73 x 10~3
-3
2.99 x 10
-3
2.72 x 10
-3
3.28 x 10
3.79 x 10~3
3.68 x 10~3
-3
3.29 x 10
-3
3.76 x 10
-3
2.94 x 10
3.46 x 10~3
-3
3.05 x 10
-3
3.43 x 10
-3
2.79 x 10
3.19 x 10~3
2.97 x 10~3
3.33 x 10~3
3.28 x 10~3
3.39 x 10~3
_3
3.24 x 10
3.87 x 10~3
-3
3.43 x 10
_3
3.72 x 10
-3
3.18 x 10
-3
3.71 x 10
3.58 x 10~3
-3
3.52 x 10
-3
3.39 x 10
3.84 x 10~3
Spleen
(g)
1.34 x 10~3
1.41 x 10~3
1.70 x 10~3
1.93 x 10~3
1.66 x 10~3
1.79 x 10~3
1.78 x 10~3
1.85 x 10~3
1.56 x 10~3
-3
1.84 x 10
-3
1.71 x 10
-3
1.95 x 10
1.53 x 10~3
1.35 x 10~3
-3
1.81 x 10
-3
2.13 x 10
-3
1.70 x 10
2.09 x 10~3
-3
1.79 x 10
-3
2.02 x 10
-3
1.56 x 10
2.12 x 10~3
1.63 x 10~3
2.07 x 10~3
1.80 x 10~3
1.81 x 10~3
-3
2./-S x 10
2.39 x 10~3
-3
2.35 x 10
-3
2.44 x 10
-3
2.27 x 10
-3
2.51 x 10
2.24 x 10~3
-3
2.41 x 10
-3
2.12 x 10
2.38 x 10~3
Rt. Kidney Ovary
(g) (paired, g)
3.99 x 10~3
3.81 x 10~3 4.60 x 10~4
3.41 x 10~3
3.41 x 10~3 3.43 x 10~4
3.12 x 10~3
3.42 x 10~3 3.31 x 10~4
3.01 x 10~3
3.48 x 10"3 3.71 x 10~4
3.04 x 10~3
-3 -4
3.34 x 10 3.70 x 10
-3
3.09 x 10
-3 -4
3.56 x 10 3.56 x 10
4.26 x 10~3
4.34 x 10~3 5.25 x 10~4
-3
3.51 x 10
-3 -4
3.75 x 10 5.21 x 10
-3
3.50 x 10.
3.92 x 10~3 3.80 x 10~A
-3
3.51 x 10
-3 -4
3.57 x 10 3.85 x 10
-3
3.37 x 10
3.78 x 10~3 3.99 x 10~4
3.60 x 10~3
3.87 x 10~3 4.40 x 10~4
4.29 x 10~3
4.21 x 10~3 5.79 x 10~4
-3
3.89 x 10
4.20 x 10~3 5.44 x 10~4
-3
3.84 x 10
-3 -4
4.14 x 10 5.33 x 10
-3
3.82 x 10
_3 -/,
3.9/ x 10 5.38 x 10
3.89 x 10~3
-3 -4
3.85 x 10 5.82 x 10
-3
3.84 x 10
3.77 x 10~3 5.40 x 10~4
Testicle
(paired, g)
2.07 x 10~2
— —
1.06 x 10~2
__
1.05 x 10~2
	
1.07 x 10~2
__
1.01 x 10~2

—
-2
1.08 x 10

—
2.06 x 10"2
	
-2
1.10 x 10

—
-2
1.14 x 10
	
-2
1.88 x 10

—
-2
1.09 x 10
_ —
1.08 x 10"2
—
2.06 x 10~2
—
-2
1.26 x 10
—
_2
1.26 x 10

—
_2
1.24 x 10

—
1.23 x 10"2

—
-2
1.17 x 10
	
                    28

-------
TABLE 11.  (Continued)
ETU
(ppm)
625
625
125
125
25
25
5
5
1
1.
0
0
625
625
125
125
25
25
5
5
1
1
0
0
525
625
125
125
25
25
5
5
1
1
0
0
Days on
Study
90
90
90
90
90
90
90
90
90
90
90
90
60
60
60
60
60
60
60
60
60
60
60
60
30
30
30
30
30
30
30
30
30
30
30
30
Sex
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M -
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
Liver
(8)
3.59 x 10"2
3.84 x 10"2
3.91 x 10"2
3.59 x 10"2
3.46 x 10"2
3.72 x 10"2
3.37 x 10"2
3.72 x 10"2
3.54 x 10"2
3.76 x 10"2
3.31 x 10~2
3.60 x 10"2
3.91 x 10"2
4.91 x 10"2
4.17 x 10"2
4.03 x 10"2
4.44 x 10"2
4.05 x 10"2
3.80 x 10"2
3.93 x 10"2
3.63 x 10"2
3.64 x 10"2
3.79 x 10~2
3.82 x 10"2
4.50 x 10~2
5.21 x 10"2
4.84 x 10"2
4.43 x 10"2
4.42 x 10~2
4.40 x 10"2
4.62 x 10"2
4.24 x 10"2
4.40 x 10"2
4.02 x 10"2
4.22 x 10"2
4.43 x 10"2
Pituitary
(g)
4.79 x 10"5
7.82 x 10"5
2.22 x 10~5
4.71 x 10"5
2.04 x 10"5
5.01 x 10"5
2.23 x 10"5
4.92 x 10*5
1.93 x 10"5
4.94 x 10"5
2.18 x 10"5
5.23 x 10"5
5.03 x 10"5
5.90 x 10"5
3.28 x 10"5
4.81 x 10"5
2.22 x 10"5
3.45 x 10"5
1.84 x 10"5
4.18 x 10"5
2.00 x 10"5
4.75 x 10"5
2.22 x 10~5
6.19 x 10"5
6.82 x 10"5
4.80 x 10"5
3.20 x 10"5
4.97 x 10"5
2.58 x 10"5
3.79 x 10"5
2.55 x 10"5
3.94 x 10"5
2.39 x 10"5
3.67 x 10"5
2.65 x 10"5
5.13 x 10"5
Thyroid
(paired; g)
22.91 x 10"5
27.10 x 10"5
7.66 x 10"5
8.73 x 10~5
5.80 x 10~5
6.88 x 10"5
5.21 x 10"5
6.55 x 10"5
5.41 x 10"5
7.14 x 10"5
5.80 x 10"5
8.81 x 10"5
5.03 x 10"5
5.90 x 10"5
3.28 x 10"5
4.81 x 10"5
2.22 x 10"5
3.45 x 10"5
1.84 x 10"5
4.18 x 10"5
2.00 x 10"5
4.75 x 10"5
2.22 x 10"5
4.19 x 10"5
15.82 x 10"5
18.90 x 10~5
10.20 x 10"5
11.20 x 10"5
7.72 x 10"5
9.89 x 10"5
5.77 x 10"5
7.93 x 10"5
6.02 x 10"5
7.06 x 10"5
6.20 x 10"5
7.14 x 10"5
Adrenal
(mg)
1.93 x 10~4
2.54 x 10~4
0.99 x 10"4
1.92 x 10"4
1.01 x 10"4
2.03 x 10"4
0.99 x 10"4
2.20 x 10"4
0.98 x 10"4
2.29 x 10"4
0.98 x 10"4
2.33 x 10"4
2.04 x 10"4
2.66 x 10"4
1.34 x 10"4
2.51 x 10"4
1.03 x 10"4
2.33 x 10"4
1.02 x 10"4
2.37 x 10"4
1.09 x 10"4
2.19 x 10"4
1.15 x 10"4
2.40 x 10"4
2.05 x 10"4
2.45 x 10"4
1.40 x 10"4
2.73 x 10"4
1.44 x 10"4
2.70 x 10"4
1.43 x 10"4
2.67 x 10"4
1.40 x 10"4
2.92 x 10"4
1.26 x 10"4
2.58 x 10"4
Brain
(8)
8.53 x 10~3
9.79 x 10"3
4.21 x 10"3
6.66 x 10"3
4.04 x 10"3
6.49 x 10~3
4.16 x 10"3
6.54 x 10"3
4.02 x 10"3
6.41 x 10"3
3.96 x 10"3
6.46 x 10"3
8.66 x 10"3
10.70 x 10"3
4.78 x 10~3
7.35 x 10"3
4.35 x 10"3
7.38 x 10~3
4.16 x 10~3
7.03 x 10~3
4.32 x 10"3
7.04 x 10"3
4.54 x 10"3
7.04 x 10"3
9.19 x 10"3
11.10 x 10"3
5.79 x 10"3
8.34 x 10"3
5.59 x 10"3
8.09 x 10"3
5.65 x 10"3
8.46 x 10"3
5.96 x 10"3
8.53 x 10"3
5.30 x 10"3
8.11 x 10"3
    29

-------
TABLE 11.   (Continued1)
Amitrole
(ppm)
50
50
50

50
50
50
PTU
(ppm)
125
125
125
125
125
125
Control
0
0
0
0

0

0
Days
on
Study
90
90
60

60
30
30


90
90
60
60
30
30

90
90
60
60

30

30
Sex
M
F
M

F
M
F


M
F
M
F
M
F

M
F
M
F

M

F
Heart
(g)
2.66
3.04
2.83

3.45
2.83
3.32


2.66
2.82
2.58
3.05
2.76
3.10

2.72
3.26
2.58
3.05

3.39

3.84
x
X
X

X
X
X


X
X
X
X
X
X

X
X
X
X

X

X
io-3
io-3
io-3
_o
10
io-3
io-3


ID'3
io-3
io-3
io-3
io"3
io-3

io-3
ID'3
io-3
ID'3
_•}
10 J
_•}
10 ^
Spleen
(g)
1.65 x
2.08 x
1.67 x

2.15 x
1.74 x
2.48 x


0.93 x
1.03 x
1.24 x
1.40 x
1.43 x
1.45 x

1.71 x
1.95 x
1.24 x
1.40 x

2.12 x

2.38 x
10"3
io-3
ID'3
-3
10 J
io-3
io-3


ID'3
io-3
io-3
io-3
io-3
io-3

io-3
io"3
io-3
io"3
_o
10
_0
10
Right
Kidney Ovary
(g) (paired) (g)
3.08
3.47
3.52

3.77
3.49
3.71


2.89
3.16
3.30
3.38
3.58
3.60

3.09
3.56
3.30
3.38

3.84

3.77
x
X
X

X
X
X


X
X
X
X
X
X

X
X
X
X

X

X
io-3
10"3 3.42 x 10"4
io-3
-3 -4
10 3.86 x 10
ID'3
10"3 6.20 x 10"4


io"3
10"3 3.08 x 10~4
ID'3
10"3 4.48 x 10"4
io-3
10~3 5.39 x 10"4

1Q-3
10~3 3.56 x 10"4
io-3
10"3 4.48 x 10"4
_o
10
-3 -4
10 6.20 x 10
Testicle
(paired) (g>
1.07 x 10"2

1.12 x 10"2

-
1.31 x 10"2



1.69 x 10"2
—
1.71 x 10"2
_
1.77 x 10"2
-

1.08 x 10"2
_
1.71 x 10"2
_
_?
1.31 x 10

—
             30

-------
TABLE 11.  (Continued)
Amitrole
(ppm)
50
50
50
50
50
50
PTU
(ppm)
125
125
125
125
125
125
Control
(ppm)
0
0
0
0
0
0
Days
on
Study
90
90
60
60
30
30


90
90
60
60
30
30


90
90
60
60
30
30
Sex
M
F
M
F
M
F


M
F
M
F
M
F


M
F
M
F
M
F
Liver
(g)
3.56 x 10~2
3.59 x 10~2
3.88 x 10~2
3.95 x 10~2
4.38 x 10~2
4.95 x 10~2


3.17 x 10~2
3.42 x 10~2
3.37 x 10~2
3.68 x 10~5
3.67 x 10~5
4.10 x 10~5


3.31 x 10~2
3.60 x 10~2
3.79 x 10~2
3.82 x 10~2
4.22 x 10~2
4.43 x 10~2
Pituitary
(g)
2.56 x 10~5
5.92 x 10~5
2.33 x 10"5
5.48 x 10~5
2.49 x 10~5
3.52 x 10~5


3.02 x 10~5
5.19 x 10~5
2.79 x 10~5
4.38 x 10~5
3.35 x 10~5
4.27 x 10~5


2.18 x 10~2
5.23 x 10~2
2.22 x 10~2
6.19 x 10~2
2.65 x 10~2
5.13 x 10~2
Thyroid
(paired) (g)
13.43 x 10~5
14.23 x 10~5
8.12 x 10~5
9.11 x 10~5
22.91 x 10~5
22.80 x 10~5


42.90 x 10~5
45.20 x 10~5
35.90 x 10~5
44.43 x 10~5
25.53 x 10~5
36.81 x 10~5


5.80 x 10~5
8.81 x 10~5
6.42 x 10~5
8.59 x 10~5
6.70 x 10~5
7.14 x 10~5
Adrenal
(mg)
0.99 x 10~4
2.09 x 10~4
1.00 x 10~4
2.52 x 10~4
1.14 x 10~5
2.44 x 10~5


1.08 x 10~4
1.40 x 10"4
1.01 x 10~4
1.68 x 10~4
1.08 x 10~4
1.64 x 10~4


0.98 x 10~4
2.33 x 10~4
1.15 x 10~4
2.40 x 10~4
1.26 x 10~4
2.58 x 10~4
       31

-------
TABLE 12.  ORGAN WEIGHT:  BRAIN WEIGHT RATIO
ETU
(ppm)

625

625

125

125
25
25
5
5

1
1

0
0

625
625

125

125

25

25
5
5
1

1
0

0
625

625
125
125
25
25
5

5

1

1

0
0
Days on
Study

90

90

90

90
90
90
90
90

90
90

90
90

60
60

60

60

60

60
60
60
60

60
60

60
30

30
30
30
30
30
30

30

30

30

30
30
Sex

M

F

M

F
M
F
M
F

M
F

M
F

M
F

M

F

M

F
M
F
M

F
M

F
M

F
M
F
M
F
M

F

M

F

M
F
Ovary
(paired; g)



4.70 x



5.15 x

5.10 x

5.67 x


5.51 x


5.28 x


4.92 x



7.08 x



5.15 x

5.48 x


5.66 x


6.25 x


5.23 x

6.52 x

6.58 x


6.36 x



6.82 x


6.69 x


_2
10


_9
10 *

io-2

io-2


io-2


io-2


io-2


_2
10 *


-2
10 i

io-2

_2
10

_2
10 ^

_9
10

io-2

ID'2

_2
10 i


-2
10 i


io-2
Testicle
(paired; g)

2.43



2.52


2.59

2.58


2.50


2.73


1.81


2.30



2.62


2.58

2.51


2.38


2.25


2.17

2.25

2.20



2.06



2.21

Pituitary
(mg)

5.61

7.99

5.27

7.08
5.04
7.72
5.36
7.52

4.79
7.71

5.50
8.09

5.81
5.54

6.85

6.54

5.09

4.68
4.41
5.95
4.63

6.75
4.89

8.80
7.21

4.34
5.53
5.96
4.63
4.68
4.52

4.66

4.01

4.31

5.01
6.36
_3
x 10
.3
x 10
.3
x 10
_3
x 10
x 10"3
x 10~3
x 10"3
x 10"3
_3
x 10
x 10"3
_3
x 10
x 10"3
.3
x 10
x 10"3
_3
x 10
_T
x 10
-3
x 10
-3
x 10
x 10"3
x 10"3
x 10"3
_3
x 10
x 10'3
-3
x 10
x 10"3
.3
x 10
x 10"3
x 10'3
x 10'3
x 10'3
x IO"3
.3
x 10 J
.3
x 10
.3
x 10
.3
x 10
x 10"3
Thyroid
(paired; g)

2.68 x

2.77 x

1.82 x

1.31 x
1.43 x
1.06 x
1.25 x
1.00 x

1.35 x
1.11 x

1.47 x
1.36 x

3.28 x
2.31 x

1.85 x

1.48 x

1.69 x

1.22 x
1.05 x
0.96 x
1.36 x

1.19 x
1.41 x

1.22 x
1.72 x

1.71 x
1.37 x
1.34 x
1.38 x
1.22 x
1.02 x

0.94 x

1.01 x

0.83 x

1.26 x
0.88 x
_9
10 *
-2
10 Z
_2
10 *
_9
10 l
ID'2
io-2
ID'2
io-2
-2
10 i
ID'2
_2
10 Z
io-2
_2
10 i
io-2
_9
10 L
_9
10 l
_9
10 l
-2
10 i
ID'2
io-2
io-2
-2
10 2
io-2
-2
10
ID'2
-2
10 i
ID'2
io-2
ID'2
io-2
io-2
_9
10 *
_9
10 i
_2
10 i
_2
10 i
io-2
Adrenal
(paired; mg)

2.26

2.60

2.35

2.88
2.49
3.13
2.38
3.37

2.44
3.51

2.47
3.61

2.36
2.49

2.79

3.41

2.38

3.16
2.44
3.37
2.52

3.11
2.53

3.44
2.23

2.21
2.42
3.27
2.57
3.42
2.52

3.15

2.36

3.43

2.38
3.19
_9
x 10
-2
x 10
-2
x 10
-2
x 10
x IO"2
x 10"2
x 10"2
x 10"2
-2
x 10
x 10"2
_2
x 10
x 10"2
_2
x 10
x 10"2
_2
x 10
-2
x 10
-2
x 10
-2
x 10
x 10~2
x 10"2
x 10"2
_2
x 10
x 10"2
-2
x 10 ^
x 10"2
-2
x 10
x 10"2
x IO"2
x 10"2
x 10"2
x 10"2
_9
x 10
_2
x 10
-2
x 10
-2
x 10
x 10"2
                   32

-------
TABLE 12.  (Continued)
(ppm)
625
625
125
125
25
25
5
5
1
1
0
0
625
625
125
125
25
25
5
5
1
1
0
0
625
625
125
125
25
25
5
5
1
1
0
0
Days on
Study
90
90
90
90
90
90
90
90
90
90
90
90
60
60
60
60
60
60
6i>
60
60
60
60
60
30
30
30
30
30
30
30
30
30
30
30
30
Sex
M
F
F
F
M
F
M
F
M
F
M
I
M
F
M
F
M
F
M
F
. M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
Heart
(g)
4.
3.
6.
4.
6.
5.
6.
4.
6.
4.
6.
5.
4.
3.
6.
5.
6.
4.
7.
4.
6.
4.
6.
4.
3.
3.
5.
4.
6.
4.
5.
4.
6.
4.
6.
4.
25 x
51 x
70 x
88 x
87 x
07 x
85 x
78 x
78 x
66 x
88 x
08 x
38 x
45 x
87 x
11 x
76 x
69 x
32 x
88 x
47 x
54 x
54 x
73 x
57 x
06 x
59 x
65 x
14 x
59 x
02 x
39 x
01 x
13 x
41 x
76 x
ID'1
lo-1
1C'1
10'1
lo-1
lo-1
lo-1
ID'1
ID'1
10'1
10'1
10"1
ID'1
10'1
10'1
10'1
lO'1
ID'1
ID'1
10'1
ID'1
ID'1
ID'1
ID'1
lo-1
ID'1
ID'1
lo-1
10'1
lO'1
lo-1
ID'1
ID'1
ID'1
lo-1
ID'1
1
1
4
2
4
2
4
2
3
2
4
3
1
1
3
2
3
2
4
2
3
3
3
2
1
1
3
2
4
3
4
2
3
2
4
2
Spleen
(g)
.57 x
.44 x
.04 x
.97 x
.10 x
.76 x
.29 x
.83 x
.89 x
.88 x
.32 x
.02 x
.76 x
.26 x
.77 x
.89 x
.91 x
.83 x
.30 x
.87 x
.61 x
.01 x
.59 x
.94 x
.96 x
.64 x
.94 x
.87 x
.21 x
.01 x
.02 x
.97 x
.76 x
.82 x
.00 x
.95 x
lo-1
ID'1
10'1
lo-1
lo-1
10'1
lo-1
ID'1
lo-1
ID'1
ID"1
lo-1
ID"1
10'1
lo-1
10'1
10'1
10'1
10'1
lO'1
lo-1
ID'1
lo-1
ID'1
ID'1
ID'1
lo-1
ID'1
ID'1
ID'1
ID'1
ID'1
lO'1
ID'1
10'1
ID'1
Rt. Kidney
(g)
4.67 x
3.89 x
8.22 x
5.12 x
7.32 x
5.27 x
7.25 x
5.32 x
7.57 x
5.21 x
7.81 x
5.51 x
4.92 x
4.07 x
7.33 x
5.09 x
8.05 x
5.31 x
8.44 x
5.08 x
7.81 x
5.37 x
7.93 x
5.50 x
4.67 x
3.81 x
6.72 x
5.03 x
6.87 x
5.11 x
6.75 x
4.70 x
6.53 x
4.51 x
7.25 x
4.67 x
10'1
lo-1
lO'1
lO'1
lo-1
lo-1
lo-1
ID'1
ID'1
10'1
lo-1
ID'1
lo-1
lo-1
ID'1
lO"1
ID'1-
10'1
10'1
10'1
10'1
10'1
lo-1
10'1
ID'1
lo-1
lo-1
ID"1
ID'1
ID'1
ID'1
ID'1
ID'1
10'1
ID'1
10'1
Liver
(g)
4.20
3.92
9.30
5.40
8.56
5.71
8.10
5.68
8.80
5.86
8.35
5.57
4.56
4.60
8.71
5 47
10.20
5.48
9.14
5.60
8.40
5.17
8.35
5.43
4.90
4.71
8.37
5.31
7.91
5.44
8.17
5.02
7.38
4.83
7.97
5.49
           33

-------
TABLE 12.  (Continued)
Amitrole
(ppm)
50
50
50
50
50
50
PTU
(ppm)
125
125
125
125
125
, 125
Control
(ppm)
0
0
0
0
0
0
Days on
Study
90
90
60
60
30
30

90
90
60
60
30
30

90
90
60
60
30
30
Sex
M
F
M
F
M
F

M
F
M
F
M
F

M
F
M
F
M
F
Heart
(g)
6.44 x 10"1
4.68 x 10"1
6.23 x 10"1
4.92 x 10"1
4.70 x 10'1 .
4.17 x 10"1

3.82 x 10'1
3.03 x 10"1
3.81 x 10"1
3.44 x 10"1
3.34 x lO'1
3.06 x 10"1

6.6>8 x 10"1
5.05 x 10"1
6.54 x 10"1
4.73 x 10'1
6.41 x 10"1
4.76 x 10'1
Spleen
(g)
3.98 x 10'1
3.20 x 10"1
3.67 x 10"1
3.06 x 10"1
2.89 x 10'1
3.12 x 10"1

1.33 x 10'1
1.10 x 10"1
1.83 x 10"1
1.58 x 10'1
1.73 x 10"1
1.43 x 10"1

4.32 x 10"1
3.02 x lO'1
3.59 x 10"1
2.94 x 10"1
4.00 x 10'1
2.95 x 10'1
Rt. Kidney
(g)
7.45 x 10'1
5.35 x 10"1
7.74 x 10"1
5.35 x 10"1
5.79 x 10"1
4.66 x 10"1

4.14 x 10"1
3.40 x ID'1
4.88 x 10"1
3.82 x 10'1
4.34 x 10"1
3.55 x 10'1

7.81 x 10"1
5.51 x 10"1
7.93 x 10"1
5.50 x 10"1
7.25 x 10'1
4.76 x 10"1
Liver
(g)
8.59
5.53
8.52
5.62
7.26
6.22

4.54
3.68
4.97
4.15
4.45
4.04

8.35
5.57
8.35
5.43
7.97
5.49

-------
TABLE 12.  (Continued)
Amitrole
(ppm)
50
50
50

50
50

50
PTU
(ppm)
125
125
125
125

125
125
Control
(ppm)

0
0

0
0
0
0
Days on
Study
90
90
60

60
30

30


90
90
60
60

30
30



90
90

60
60
30
30
Ovary
Sex (paired; g)
M
F 5.28 x 10~2
M
-2
F 5.50 x 10
M
_2
F 7.79 x 10


M
F 4.17 x 10~2
M
F 5.06 x 10~2

M
F 5.32 x 10"2



M
F 5.51 x 10"2

M
F 6.21 x 10~2
M
F 6.69 x 10"2
. Testicle
(paired; g)
2.58
—
2.47

—
2.17




2.42
._ —
2.52
—

2.14
—



2.73
—

2.38
—
2.21
	
Pituitary
(mg)
6.20
9.13
5.12

7.79
4.13

4.42


4.39
5.58
4.11
4.94

4.05
4.21



5.50
8.09

4.89
8.80
5.01
6.36
x
X
X

X
X

X


X
X
X
X

X
X



X
X

X
X
X
X
io-3
io-3
io"3
_3
10 J
io"3
_T
10 J


io-3
io-3
io"3
io-3
_•}
10 J
10"3


_•}
10 J
io-3
_•}
10 J
io-3
io-3
io"3
Thyroid
(paired; g)
3.23
2.20
1.79

1.30
3.80

3.13


6.14
4.86
5.30
5.01

3.09
3.64



1.47
1.36

1.41
1.22
1.26
0.89
x
x
X

X
X

X


X
X
X
X

X
X



X
X

X
X
X
X
io-2
io-2
io"2
-2
10
io-2
_2
10 Z


io-2
io-2
io"2
io-2
_2
10 Z
io-2


_?
10 L
io-2
_o
10 ^
io-2
io-2
io-2
Adrenal
(paired; mg)
2.40 x
3.22 x
2.20 x

3.59 x
1.88 x

3.67 x


1.55 x
1.50 x
1.49 x
1.90 x

1.30 x
1.62 x



2.47 x
3.61 x

2.53 x
3.41 x
2.38 x
3.19 x
10-2
io-2
io"2
_o
10
io-2
_2
10


io-2
io"2
io-2
io"2
_ 0
10
io-2


— 7
10
io2
— 7
i fl-
ic"2
io-2
io-2

-------
                     TABLE  13. ABNORMAL CHANGES OBSERVED FOR LIVER, SKIN AND THYROID

Tissue Change
Enlarged Thyroids; Red

Thyroids Normal Size but Red

Centrilobular Congestion; liver

Alopecia

Small Adrenals

Enlarged Thyroids; Red

Centrilobular Congestion; liver

Alopecia

Days on
Study
30
30
30
30
30
30
30
30
30
30
60
60
60
60
60
60
Sex
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
PTU Amitrole
125 50
10
10
0
0
0
0
0
0
0
0
10
10
0
0
0
0
10
10
0
0
0
0
0
0
8
2
5
2
2
0
0
0
ETU
625
10
5
0
0
0
0
8
4
0
0
5***
9
0
0
5
9
125
8
2
0
0
0
0
0
0
0
0
6
1
2
0
0
0
25
0
0
3
2
0
0
0
0
0
0
1
0
3
0
0
0
5
0
0
0
0
0
0
0
0
0
0
2
2
2
. 0
0
0
Control**
1
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1
0
0
0
****
Enlarged Thyroids; Red

Alopecia

Centrilobular Congestion; liver

Small Adrenals

Adenocarcinoma of Salivary Gland

90
90
90
90
90
90
90
90
90
90
M
F
M
F
M
F
M
F
M
F
10
10
0
0
0
0
8
9
0
1
10
5
0
0
3
2
0
0
0
0
8
6
6
4
0
0
0
0
0
0
10
6
0
0
9
0
0
0
0
0
0
0
0
0
1
1
0
0
0
0
0
0
0
0
3
0
0
0
0
0
0
0
0
0
6
2
0
0
0
0
0
0
0
0
1
0
0
0
0
0

   *Presented as the number of animals with  the  indicated  tissue change,  out of the 10 animals evaluated.

  **Twelve male and eleven female control  rats were  evaluated  from  30  day group.   Twelve male and twelve
    female rats were evaluated from the 60 and 90 day control  groups.

 ***Five *male rats and one female rat  from the 625 ppm 60  day  group died  prior to the end of the study.
    Therefore, the values for this one group are based on  the  evaluation  of  five male and one female rat.

****Two female rats died prior to the  end  of the 90  days.   Therefore,  the values for females on this one
    group  are based on eight rats.

-------
TABLE  14.  GRADING OF DEGREE OF HYPERPLASIA FROM MICROSCOPIC
           APPEARANCE OF THE THYROID GLAND
  Grade of
Hyperplasia	Description of Change in Morphology	

    0             Completely normal appearing gland.
   >1             Very slightly altered lining cells and
                    scalloping of colloid.
    1             Distinct hyperplasia or hypertrophy
                    of lining cells and a moderate
                    reduction of colloid.
   >2             Hyperplastic change of lining cells
                    and folding of follicular wall with
                    a moderate reduction of colloid.
    2             Moderate hyperplasia of lining cells,
                    increase in apparent number of micro-
                    follicular follicles reducing apparent
                    colloid to less than fifty percent of
                    normal.
   >3             More extensive microfollicular pattern
                    but still retaining approximately
                    twenty-five percent colloid.
    3             Marked hyperplasia of lining cells with
                    virtually all microfollicular follicles
                    and nearly complete absence of colloid.
                              37

-------
TABLE 15.THYROID HYPERPLASIA OBSERVED IN ETU TEST ANIMALS

Degree of
Hyperplasia
0
.>!
1
>2
2
>3
3
0
>1
1
>2
2
>3
3
0
>1
1
>2
2
>3
3
Days on
Study
30
30
30
30
30
30
30
60
60
60
60
60
60
60
90
90
90
90
90
90
90
PTU
125
0
0
0
0
8
3
9
0
0
0
0
0
17
3
0
0
0
0
1
10
9
Amitrole
50
0
0
0
. 0
0
5
15
0
4
5
8
3
0
0
0
5
14
0
1
0
0
625
0
0
2
4
12
2
0
0
0
2
2
10
0
0
0
0
1
1
12
3
1
125
0
0
18
1
1
0
0
1
3
7
5
2
0
2
1
0
14
3
1
0
1
ETU
25
0
9
11
0
0
0
0
7
2
0
7
4
0
0
3
8
9
0
0
0
0
5
1
6
13
0
0
0
0
4
7
9
0
0
0
0
6
8
6
0
0
0
0
1
0
7
13
0
0
0
0
1
9
10
0
0
0
0
6
6
8
0
0
0
0
0
4
9
10
0
0
0
0
5
10
9
0
0
0
0
12
9
3
0
0
0
0
                            38

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    TABLE 16.  CHANGES IN THYROID MORPHOLOGY IN RATS WHICH
               INGESTED 625 PPM ETU FOR NINETY DAYS*

Rat
Number
560
561
562
563
564
565
566
567
568
569
570
571
572
573
574
575
576
577
578
579
Sex
M
M
M
M
M
M
M
M
M
M
F
F
F
F
F
F
F
F
F
F
Solid
Adenomas
1
1
1
0
1
0
0
2
0
0
Died
Died
2
0
0
0
0
0
1
0
Cystic
Adenomas
0
2
1
0
0
0
0
0
0
0


0
0
0
0
0
0
0
0
Adenomatous
Changes
0
2
0
1
2
0
0
0
0
0


1
0
0
1
0
0
0
0
Cyst-like
Follicles
0
0
0
0
0
1
0
0
1
0


0
1
0
0
0
1
0
0

*
 Thyroids examined by serial sectioning.
                             39

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                                  TECHNICAL REPORT DATA
                           (Please read Instructions on the reverse before completing)
i. REPORT NO.
  EPA-600/1-77-023
                                                          3. RECIPIENT'S ACCESSION NO.  .
4. TITLE AND SUBTITLE

  DIETARY SUBACTURE TOXICITY  OF  ETHYLENE THIOUREA IN THE
  LABORATORY RAT
                 5. REPORT DATE
                    May  1977
                 6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
                                                          8. PERFORMING ORGANIZATION REPORT NO.
  Ralph I. Freudenthal
9. PERFORMING ORGANIZATION NAME AND ADDRESS
  Battelle Columbus Laboratories
  505 King Avenue
  Columbus, Ohio 43201
                                                           10. PROGRAM ELEMENT NO.
                    1EA615
                  11. CONTRACT/GRANT NO.

                   68-02-1715
12. SPONSORING AGENCY NAME AND ADDRESS
  Health Effects  Research  Laboratory
  Office of Research and Development
  U.S.  Environmental Protection Agency
  Research Triangle Park,  N.C. 27711
                                                           13. TYPE OF REPORT AND PERIOD COVERED
HERL-RTP
                  14. SPONSORING AGENCY CODE
                    EPA  600/11
15. SUPPLEMENTARY NOTES
16. ABSTRACT
  Ethylene thiourea  (ETU) was  fed to groups of rats at 0,  1,  5,  25,  125 or 625 ppm for
  up to 90 days.  Other  groups of rats received either propulthiouracil (PTU; 125 ppm)
  or amitrole (50 ppm) in their diets as positive controls. Only those rats which
  received ETU at 125 or 625  ppm and those ingesting PTU or amitrole demonstrated a
  measurable toxic response.   This toxicity was reflected  as  an  alteration in thyroid
  function and a significant  change in thyroid morphology.     Ingestion of 625 ppm ETU
  or 125 ppm PTU resulted in  very substantial decreases in serum triiodothyronine (T-3)
  and thyroxine (T-4). Marked  increases in serum thryroid  stimulating hormone (TSH)
  levels were found  in the  625 and 125 ppm ETU rats, the 125  PTU rats and the rats
  receiving amitrole, at each  time point this hormone was measured.   A decrease in
  iodide uptake by the thyroid was also found in the rats  ingesting  625 ppm ETU.
  Wnile a statistically  significant increase in serum T-4  and  degree of thyroid hyper-
  plasia was observed for the  rats ingesting 25 ppm ETU for 60 days, normal thyroid
  normone levels and thyroid morphology was found in the rats  on 25  ppm ETU for either
  JO or 90 days.     Based on biochemical and microscopic changes examined, the no-effect
  level for dietary  ETU  in  this 90-day study is considered to  be 25  ppm,  equivalent to
  an average intake  ranging from 19.5 mg/kg body weight at Week  1  to 12.5 mq/kq body
  weight at Week 12.                                                .         3/  y    y
17.
                               KEY WORDS AND DOCUMENT ANALYSIS
                  DESCRIPTORS
    b.IDENTIFIERS/OPEN ENDED TERMS  C.  COSATI Field/Group
  thioureas
  toxicity
  toxic  tolerances
  rats
     ethylene thiourea
     ETU
06, T
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                                            40

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