United States
               Environmental Protection
               Agency
                 Health Effects Research
                 Laboratory
                 Research Triangle Park NC 27711
FPA-600 1 78-056
August 1978
               Research and Development
EPA
Teratology  of
Guthion
                          LI

-------
                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
      5.  Socioeconomic Environmental Studies
      6.  Scientific and Technical Assessment Reports (STAR)
      7.  Interagency Energy-Environment Research and Development
      8.  "Special" Reports
      9.  Miscellaneous Reports
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
studies. In addition to toxicology and other medical specialities, study areas in-
clude biomedical  instrumentation and health research techniques utilizing ani-
mals — but always with  intended application to human health measures.
This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia 22161.

-------
                                       EPA-600/1-78-056
                                       August 1978
             TERATOLOGY OF GUTHION
                      by

Robert D. Short, Jan L. Minor, Timothy M. Unger
              and Cheng-Chun Lee
          Midwest Research Institute
               425 Volker Blvd.
            Kansas City, Mo. 64110
            Contract No. 68-02-2746
                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

-------
                            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

-------
                                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 tcxicology,
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 participates  in
the development and revision of 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 primarily responsible for providing
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 study described in this report was conducted under our program
to determine the health implications of substances used as pesticides, in
an effort to evaluate the teratogenic potential of Guthion.
                                   F. G. Hueter, Ph. D.
                                     Acting Director,
                           Health Effects Research Laboratory
                                   iii

-------
                           TABLE OF CONTENTS

                                                                      Page

Summary 	   1

I.      Introduction 	   2

II.    Methods	   2

         A.   Animals	   2
         B.   Guthion and Preparation of Dose	   2
         C.   Toxicity	   3
         D.   Teratology	   3

III.   Results	   4

         A.   Toxicity Study 	   4
         B.   Teratology Study in Rats and Mice Treated with Guthion  .   5

               1.  Maternal Welfare and Reproduction	   5
               2.  External Anomalies . 	   5
               3.  Soft Tissue Anomalies	   5
               4.  Skeletal Anomalies 	   6

         C.   Toxicity of Guthion in Rats Treated During Gestation and
               Lactation	   6

IV.    Discussion	   7

References	   9

Tables 1-12	10-21
                                    IV

-------
                                SUMMARY
          The purpose of this study was to assess the effects of Guthion,
a pesticide with anticholinesterase activity, on development in rats and
mice.  A preliminary toxicity study with Guthion indicated that a 35 day
LD5Q dose for virgin rats and a 10 day LDcn dose for virgin mice was be-
tween 4 and 8 mg/kg/day for both species.  On the basis of this data, doses
of 0, 1.25, 2.5 and 5.0 mg/kg/day were selected for the developmental study,
which consisted of two phases.  During the first phase, pregnant rats and
mice were treated for 10 days starting on gestational day 6.  The high
dose affected maternal welfare only in rats.  Guthion did not significantly
increase in a dose-related manner any of the specific anomalies observed in
either rats or mice.  During the second phase, pregnant rats were treated
from gestational day 6 to postpartum day 21.  Dams in the high dose group
were more sensitive to Guthion later in gestation with the result that
deaths and signs of anticholinesterase toxicity increased during this time.
Guthion also adversely affected maternal welfare in this group.  As a re-
sult of Guthion toxicity, only one litter survived until weaning.  The
inability to dissociate toxicity in adult and developing animals suggests
that Guthion has little primary effect on the development of rats or mice.

-------
                         TERATOLOGY OF GUTHION
                           I.  INTRODUCTION
          The purpose of this study was to evaluate the teratogenic poten-
tial of Guthion, a pesticide with anticholinesterase activity.  The study
consisted of two phases.  The first phase was a toxicity study and involved
treating virgin female rats and. mice with Guthion.  The purpose of this
phase was to establish doses for the second phase.  During the second phase,
pregnant rats and mice were treated during gestation with Guthion and their
fetuses were examined for abnormalities.  In addition, some pregnant rats
were treated during gestation and lactation and their pups were observed
until weaning.
                             II.  METHODS
A.  Animals

          Charles River CD® rats weighing 200 to 220 g and CD®-1 mice weigh-
ing 20 to 22 g (Charles River Breeding Laboratories, North Wilmington,
Massachusetts) were housed in our animal quarters for at least 7 days prior
to use.  The quarters were maintained at 22 to 26°C with a relative humidity
of 40 to 60% and a 7:00 AM to 7:00 PM photoperiod.   Animals were given free
access to tap water and rodent chow  (Wayne Lab-Blox, Allied Mills, Inc.,
Chicago, Illinois).

          Pregnant animals for the teratology study were mated in our animal
quarters.  Females were housed overnight with a male of the same species.
The following morning, rats were examined for sperm in vaginal smears and
mice were examined for vaginal copulatory plugs.  The presence of these  signs
was taken as evidence of successful mating, which was identified as day  0
of gestation.
B.  Guthion and Preparation of Dose

          A sample of Guthion  (Azinphosmethyl, Lot No. M007, technical  grade),
which was received from Environmental Protection Agency  (Research Triangle
Park) on September 22, 1977, was used in this study.  The Guthion was admin-
istered orally in a vehicle of cold pressed corn oil  (Hain  Pure Food Co.,
Los Angeles, California) which was reported to be free of preservatives.
This vehicle was selected to prevent a possible interaction between preserva-
tives and Guthion.  All doses were administered in. a volume of 5 ml/kg  of
corn oil.

-------
          The high dose was prepared by dissolving a weighed amount of
Guthion in acetone, adding the calculated volume of corn oil, and heating
the solution for 20 min at 65 to 75°C to volatilize the acetone.  Lower
doses were prepared by serial dilutions of the high dose.  The volume of
acetone prior to heating was calculated to represent 2% of the final volume
of corn oil.  The control group received a solution of corn oil and 2%
acetone which was heated for 20 min at 65 to 75°C.  All solutions were pre-
pared fresh daily and administered at room temperature.
C.  Toxicity

          Virgin female rats and mice were treated with daily oral doses
of 0, 0.5, 1.0, 2.0, 4.0, 8.0 and 16.0 mg/kg of Guthion.  The body weight
on the first day of treatment was used for calculating all doses.  Rats
were treated for 35 days and mice were treated for 10 days.  All animals
were observed for 10 additional days at the end of the treatment period.
D.  Teratology

          Pregnant rats and mice were treated with daily oral doses of
0, 1.25, 2.5 and 5.0 mg/kg of Guthion.  The body weight on day 6 of ges-
tation was used for calculating all doses.  For rats, each group was
divided into two sub-groups.  The first sub-group was treated from day 6 to
15 of gestation and sacrificed on day 20 of gestation for examination of the
fetuses.  The second sub-group was treated from day 6 of gestation until
the pups were weaned, 21 days after birth.  For mice, dams were treated
from day 6 to 15 of gestation and sacrificed on day 18 of gestation for
examination of the fetuses.

          For the fetal examinations, pregnant animals were sacrificed by
cervical dislocation and a laparotomy was performed.  The number of live,
dead and resorbed fetuses was determined.  The umbilical cord was clamped
and severed distally to prevent blood loss.  Live fetuses were removed,
weighed, and.immediately examined for external anomalies.  One-half of the
viable  fetuses from each litter was fixed in Bouins fluid for 2 weeks.  The
hardened fetuses were serially sectioned from  the head through the trunk into
1-mm thick sections and examined for soft-tissue anomalies. —'  The remain-
ing fetuses were fixed in  70% alcohol for 2 weeks and eviscerated.  The
 fetuses were  stored  in  1% KOH  for  2 days  and  then stained with  alizarin
 red.!/  Af
 anomalies.
    2/
red.—   After differential decolorization, the skeletons were examined for

-------
E.  Statistics

                                                                3/
          Data were analyzed for homogeneity by Bartlett's test.—   Homo-
geneous data were analyzed by Dunnett's procedure.—-   Heterogeneous data
were analyzed by a nonparametric rank test.—'  The level of significance
was selected as p<0.05 unless otherwise indicated.  The litter was con-
sidered the experimental unit of observation.  For example, the percent of
fetuses with a given anomaly was calculated for each litter and these values
were averaged and expressed as the mean + standard error (S.E.) for a treat-
ment group.  Therefore, the mean value provides a measure of the affected
fetuses per litter, and the standard error provides an estimation of the
distribution of effect between litters within a group.  Data are reported
as the mean + S. E.
                              III.  RESULTS
A.  Toxicity Study

          The mortality in rats treated with Guthion for 35 days and mice
treated with Guthion for 10 days is presented in Table 1.  All rats treated
with 8 or 16 mg/kg/day of Guthion died within 5 or 2 days, respectively.
Deaths which occurred at lower doses were attributed to injury produced by
dosing as evidenced by tearing of the esophagus.  Therefore, deaths which
were due to Guthion toxicity occurred only in rats treated with 8 or 16
mg/kg/day.  Other signs of toxicity included salivation, urination, lacri-
mation and tremors.  These signs were observed after treatment with 8 and
16 mg/kg/day and occurred within an hour of dosing.  Rats exhibited these
signs of toxicity for several hours and appeared to be asymptomatic the
next day.  Similar signs of toxicity were not evident during the 35-day
treatment period with lower doses.  No signs of toxicity were observed in
any of the surviving rats during the 10-day recovery period.

          In mice, similar effects were observed (Table 1).  All deaths,
which were attributed to Guthion toxicity, occurred in the groups that re-
ceived 8 or 16 mg/kg/day.  Other signs of toxicity in these groups included
salivation, urination, lacrimation and tremors.  The duration of these signs
in mice was similar to the duration in rats.  No signs of toxicity were
observed in mice treated with doses of 4.0 mg/kg/day or lower of Guthion
or in surviving mice during the 10-day recovery period.

-------
B.  Teratology Study in Rats and Mice Treated with Guthion

          1.   Maternal Welfare and Reproduction

               a.  Rats:  Adverse effects on maternal welfare were observed
only in rats that received 5 mg/kg/day of Guthion (Table 2).  These effects
included reduced weight gain and feed consumption during the treatment
period.  These changes were reversible since the values returned to normal
when treatment was terminated.  Signs of anticholinesterase toxicity, which
included tremors, salivation, and urination, occurred in a few rats from
this group.  Although one death was observed in this group, the data are
insufficient to evaluate the effect of pregnancy on toxicity.  The remaining
groups of rats appeared to be normal.  In addition, treatment did not sig-
nificantly affect the litter size, incidence of resorptions, or fetal body
weight in any of the groups tested.

               b.  Mice:  Guthion did not affect the maternal weight change
or feed consumption at any of the doses tested (Table 3).  Mice and rats
demonstrated similar signs of anticholinesterase toxicity to 5 mg/kg/day of
Guthion; however, the incidence of these signs was lower in mice.  Although
one death was observed in the high dose group, no conclusions can be made
concerning the effect of pregnancy on toxicity.  Guthion, in addition, did
not alter the litter size, incidence of resorptions, or fetal body weight
in any of the groups tested.

          2.  External Anomalies

               a.  Rats:  Umbilical hernia and hematoma on the left foot
were observed in single fetuses from different litters in the group that
received 1.25 mg/kg/day of Guthion.  Hematoma on the trunk was also observed
in a single fetus from the group that received 2.5 mg/kg/day of Guthion.
No external anomalies were observed in any of the other groups.

               b.  Mice:  No external anomalies were observed.

          3.  Soft Tissue Anomalies

               a. Introduction:  The soft tissue anomalies observed in rats
and mice are summarized and ranked in Table 4.  All anomalies were assigned
a rank which reflects our opinion of their value in predicting teratogenic
potential.  According to this ranking, anomalies with a rank of 1 are con-
sidered to have little value in predicting teratogenic potential.  These
anomalies may occur spontaneously or represent an artifact of preparation.
In contrast, anomalies with a rank of 3 occur infrequentlv in controls and
are indicative of a disruption in normal development which m^v Compromise
survival.  Anomalies with a rank of 2 are thought to b--  •• "  . -•:crriedj ate value
in predicting teratogenic potential.

-------
               b.  Rats:  These anomalies are summarized and ranked in
Table 5.  Guthion treatment at doses of 1.25 mg/kg/day was associated with
an increase in slight lateral and fourth ventricle hydrocephalus.  The in-
cidence of this anomaly was not related to the dose.  The summary of anoma-
lies by rank indicated that Guthion did not significantly increase the
incidence of anomalies which were felt to be predictive of teratogenic
potential.

               c.  Mice:  These anomalies are summarized and ranked in Table
6.  Guthion treatment was not associated with a statistically significant
increase in any of the reported anomalies.  When the anomalies were sum-
marized by rank, there was a significant increase in the combined anomalies
with ranks of 2 and 3 in the groups treated with 2.5 and 5.3 mg/kg/day of
Guthion.  In addition, anomalies with a rank of 3 were observed only in the
groups treated with Guthion.

          4.  Skeletal Anomalies

               a.  Introduction:  The anomalies observed in rats and mice
are summarized and ranked in Table 7.  The previously described criteria
was used in assigning a rank to individual anomalies.

               b.  Rats:  These anomalies are summarized and ranked in
Table 8.  Guthion did not significantly increase in a dose-related fashion
any of the skeletal anomalies observed in rats.

               c-  Mice:  These anomalies are summarized and ranked in
Table 9.  Guthion significantly increased the incidence of malaligned
sternebrae in the group that received 5.0 mg/kg/day.  The anomalies with a
rank of 2 were also increased in this group.


C.  Toxicity of Guthion in Rats Treated During Gestation and Lactation

          Guthion produced death, and reduced both  the feed, consumption and
weight gain of dams treated with 5.0 mg/kg/day from gestational day 6 until
postpartum day 21 (Table 10).  Most of these deaths occurred prior to delivery.
In addition, the incidence of dams demonstrating anticholiriesterase toxicity
increased prior to birth.  None of the other doses  produced death or reduced
the feed consumption  and weight gain of dams similarly treated.  The duration
of gestation ranged from 21 to 23 days.

          Pup weight  (Table 10) was adversely affected by 5.0 but not 1.25
or 2.5 mg/kg/day of Guthion.  The percent of pups suriviving at various
intervals was reduced in the group treated with the high dose of Guthion
(Table  11).  None of  the treatments significantly altered the fertility or
gestation index  (Table 12).  However, there was a trend towards a reduced
gestation index in the high dose group.

-------
          Neuromuscular problems in the offspring were observed in this
study.  One day after weaning, pups in the single surviving litter of the
high dose group were observed to (1) maintain their rear legs at right
angles to the body and have muscular incoordination in the use of these
legs, (2) have muscle tremors in the tail, and (3) have an upturned snout.
In this litter of five pups, these effects were noticeable in two pups and
of questionable incidence in two.  Similar symptoms were also observed in
one pup from the control group.  Therefore, attempts to correlate these
neuromuscular problems with Guthion treatment were complicated.

          Pups were sacrificed at 30 to 40 days of age and preserved in
neutral buffered formalin.  There were 25 males and 25 females from 13 litters
in the control group, 23 males from 11 liters and 20 females from 10 litters
in the group treated with 2.5 mg/kg/day of Guthion, and 1 male and 1 female
from the group treated with 5.0 mg/kg/day of Guthion.  No offspring were
saved from the group that received 1.25 mg/kg/day of Guthion.
                            IV.  DISCUSSION

          Although an acute oral LDrg dose could not be calculated, this
value is probably between 4 and 8 mg/kg/day for both mice and rats.  The
majority of deaths in both species occurred within 5 days from the start
of treatment.  Other signs of toxicity included salivation, urination,
lacrimation and tremors.  These symptoms were observed within an hour of
dosing with both 8 and 16 mg/kg/day of Guthion and lasted for several hours.
Similar signs of toxicity were not observed with lower doses of Guthion in
either rats or mice.  No signs of delayed toxicity occurred in either species
during the treatment or recovery period.  On the basis of these results,
doses of 0, 1.25, 2.5 and 5.0 mg/kg/day of Guthion were selected for the
developmental toxicity studies.

          Adverse effects on maternal welfare in rats were observed only in
the group that received 5.0 mg/kg/day of Guthion.  These effects were a reduced
weight gain and feed consumption during the treatment period.  Other signs of
toxicity in this group included salivation, urination, lacrimation and
tremors.  In mice, similar parameters of maternal welfare were not signi-
ficantly affected at any of the doses tested.  Mice also responded to 5.0 mg/
kg/day of Guthion with salivation, urination, lacrimation and tremors; however,
the incidence appeared to be lower.  The data from these studies is insuffi-
cient to determine if pregnancy affects the toxicity of Guthion.

          Guthion, administered for 10 days during gestation, did not affect
the litter size, incidence of resorptions, or fetal body weight in either rats
or mice.  In rats, Guthion did not produce either a dose-related increase
in any of the observed anomalies or a characteristic pattern of anomalies.

-------
In mice, none of the individual anomalies increased in a dose-related fashion
or produced a characteristic pattern of defects.  However,  when all of the
anomalies were combined by rank there was a significant increase in soft-tissue
anomalies with a rank of 2 and 3 in the group that received 2.5 and 5.0 mg/kg/
day of Guthion.  In addition, skeletal anomalies with a rank of 2 were in-
creased in the group that received 5.0 mg/kg/day of Guthion.  Therefore, in
mice the combined incidence of anomalies with an intermediate value in pre-
dicting teratogenesis was increased; however, there was not a characteristic
pattern of defects.  This observation suggests that Guthion increased the
incidence of naturally occurring anomalies in mice without  producing specific
defects.

          Additional groups of rats were treated with Guthion from gestational
day 6 until postpartum day 21.  This treatment schedule produced more deaths
in dams receiving 5 mg/kg/day of Guthion than the previous   10-day treatment
schedule.  Most of these deaths occurred prior to delivery  and the incidence
of rats demonstrating anticholinesterase toxicity also increased during this
time.  These observations indicated that dams were more sensitive to Guthion
later in gestation.

          In addition to mortality, treatment during gestation and lactation
reduced feed consumption and weight gain of dams treated with 5.0 but not
1.25 or 2.5 mg/kg/day of Guthion.  The high dose also reduced the survival
and weight gain of pups from these dams.  In contrast, neither 1.25 nor 2.5
mg/kg/day of Guthion altered any of the above parameters.

          In order to be classified as a teratogen an agent must alter  the
structure or function of a statistically significant number of young.—
An agent is not classified as a teratogen if it only produced fetal death or
reduces fetal  growth.  In addition, an agent is not classified as a teratogen
if the dose required to produce an effect in the embryo or  fetuses is overtly
toxic to the dam.  Therefore, the  inability  to dissociate maternal and  fetal
effects in the present study suggests that Guthion has little primary effect
on development and, according to the above criteria, is not a teratogen in
either  rats or mice.

-------
                              REFERENCES
1.   Wilson,  J.  G. ,  "Methods for Administering Agents and Detecting Malfor-
      mations in Experimental Animals," In Teratology—Principles and
      Techniques,  J.  G.  Wilson and J. Workany (eds.)> University of Chicago
      Press, Chicago, Illinois, pp. 262-277 (1965).

2.   Staples, R.  C.  and V. L. Schnell, "Refinements in Rapid Clearing Tech-
      niques in the KOH-Alizarin Red S Method for Fetal Bones," Stain
      Technol.,  39:61-63 (1964).

3.   Steel,  R. G. D. and J.  H. Torrie, Principles and Procedures of Statistics,
      McGraw-Hill Book Co., New York (1960).

4.   Mann, H. B.  and D. R. Whitney, "On a Test of Whether One of Two Random
      Variables is Stochastically Larger than the Other," Ann. Math. Stat.,
      18,: 50-60 (1947).

5.   Staples, R.  E.  and J. G. Wilson, Definition of teratogenesis and tera-
      togen, in Methods for Detection of Environmental Agents that Produce
      Confenital Defects (T. H. Shepard and J. R. Millers, eds.) pp. 25-26,
      American Elsevier Publishing Company, New York (1975).

-------
                                TABLE 1
        MORTALITY IN FEMALE RATS AND MICE TREATED WITH GUTHION
Dose
(mg/kg/day)
0
0.5
1.0
2.0
4.0
8.0
16.0

Dose
(mg/kg/day)
0
0.5
1.0
2.0
4.0
8.0
16.0
Actual
Number
Treated Dead
6 0
6 IS/
; $
6 0
6 6
6 6

Actual
Number
Treated Dead
6 0
6 0
6 0
7 1-'
6 0
6 6
6 6
Rats
Deaths on Corrected
Treatment Day Number^/
1 2 3 4 5 13 35 Treated Dead
6 0
1 50
1 60
1 60
6 0
312 66
15 66
Mice
Deaths on Corrected
Treatment Day Number—
12345 9 10 Treated Dead
6 0
6 0
6 0
1 60
6 0
113 1 6 6
6 66
_a/  Death attributed to dosing rather than Guthion.
b/  Number of animals that died from dosing were subtracted.
                                     10

-------
x-v ^ ~^ -^. -~~~ CT>
i— i o oo ^o m m tn i— ii— i o 
"3.
I
c
o
*r*4
x;
3
C^












°. r-4 c^ o' oo ^ +1 +1 +1 +1+1 r^ +1 +1 0 +1
U-1 CM 1— 1 r-l
ON O^ OO O^ ON 00 ^3 ^O r
CM in -J- r-4 CN -ON
CM
r-l

^ ^ ^ ^ "*.
mOO^O encMCn r-lr-l Or-4 r-l
0 r-l
-1 r-4' 0
+•1 0 r^ +| +1
I"^
—4 iO ON
• -
r-4 en

*«^ r^
o' o'
"^ r-4 r^" o" S' O' +| +| +| +1+1 0 +| +| 0 +1 0 0 0+| +1
CM CM CN CN
>ocMr~- -d-a\ CTS«* in
^o ^o ^o ^**J ^**J • fy\
en


s-\ s~*> m
f— ^ ^,1^ f— ^ x-^ •
i— i o cr> O enencn i— < r-i OCN CM
CM r+ CM" o^ a^ o1 -H +| +| +1+1 ON +1 +| o +1
CM r-l --4
r-4 r-*Or—i moo -v •
1-40000 cnen^t I-ICM OCN CM
Ol r-4 5" o' r^ o' +1 +1 +! +1+1 i*- +1 +1 o +1
CN r-l r-l
rHcMr-i moo CM-* m
\O vO f*^ CM CM • ON
r-l











CO
C
O
-^*. 73 *r-4
Oj| •'-^ (U 4J
s*-\ Q) 4J qj 1 •^^^ 4J CO CO 03 0<
5< &C C -u^l C U J-t D  cfl CO r<
4-1 CO >-x — ' X-N CUcOCl) cjl 4) 0} Vt -O fl» M-i
CO C S-S f-S 4J V4 -i C! M
(-1 1 UJ 30 0) 3-1-4C003 1— 1 i-l CO CO i— 1
CUCQCOQ ^4Jg M4J cas^s-^s^
^50 M ^34-icO TD3^ 6CE
gZ P< T3Q> £
CO 3
-Ho ON +! +1 -a -o
1-4 4)
r-4 m oo c a
• • 0 0
CM en M
r-4 4J a
,(**;
oo TO
1-1
CD 4J
3 4J
in r-l fll
• • 4) C
r-4 O O C fi
+|0 r-. +| +| l! (S
r-l 0) 	 '
f— i *^ cy\ 4J
3 r-4
en en o
r-4 1 W
4J
0s o
a
^s,
co a) g
c -no
0 )-c
J 1 t\f\
4J r-4
OL. CO O 4J
r4 > CM C
O rJ CO
03 V >, • )J
, 0)
CO W C 13 CO 4-1
C i-i ^-^ TJ ^w
O CO -^ ^. 1-4
i-l 4J x-v 4-J 4-1 4-J •O
4-J CO g CO JT 03
O. r-l 60 i-l 60 ^1 ^
I-l CX ~^ i-l r-4
O g 4J CO 4J 4-1
03O 4J CSCC
CO cj g rC co co CO
I-i cocObO C^CO
£ M"Oi-l 60T360i-l
CO 4J CO ^* CO CO O CO M-4
4-11-4 4JC03 V4J2I-I1-I
cfl 3 4-J  Cr, fc
-r-^ ^s^ -".^ 	 •x.v_ «N^^
f-3 «3| ^| ul t3l
11

-------
en

y-v /— v oo in in CN -J-
CM /—- CO y-x ... .
enOvor^ OOO OO








,— ^
i.
2
O

C/3
h-l
CO
W
td
a
o
2

O
oi
O

O
2
M
oi

Q

2
O
i— i
H

H

M
2
M
S
a
**•
2
O
r—
pr
E;
C

O

H

EL
fa
fa
W
cd
*o
^^
61

^s.
W
w
o
M
^

2


2
O
M
H
U
p
Q
O
Pi
CM
Cd
oi

Q
2


U
ai

fa
Ed
^

j_j
^
2
£
H
38
c

C
0
T-l
r*|
4J
3
O













°. CN X o" in" ^ +1 +1 +1 +1+1
m CM 1—4
*^ ^o os *^5 cn
... • .
in en CM ^o m
.-i

y— v y* N in en *^* i-^ en
en y—^ r*^ • E
CUC 5-« U CO 4-1 G (0 -u
4-» CO y^ ^ '-N 01 co 0) Olcd
co c s~s 5-5 4Jwcuvi cui-ioi
0) 60 •— ' 4J'~' ,£ 4-1 )-i i-4 ^ 01 3 co O g
T-4 g <4-4 4-1 0) CO O
> co CU l-i CU u
>j T3  Q W rJ T-I
03 1-4 CO rJ
c a- 5^ s-5 f^ s~5 e
6£ 6 to 01
0) M Q >
V-l t-4
PH J
en
\»o c^
. .
0 0
+1 +1

en r^
• en
OS
1— 1
CN


p>^
CO
• 13
r— i
•>
(-1 00
01 r-4
c ?i cd
i-i cd -a
-^ T3 -^
01 •*-' 01
co ca
3 4J 3
E "60 g
T-4
4J O) .u
C S C
cd co
C >, c
60 T3 60
0) 0 \ (J\
                                                                             12

-------
                           TABLE 4
      SUMMARY OF OBSERVED SOFT TISSUE ANOMALIES BY RANK
Rank 3:  Most valuable in assessing teratogenic potential.

             Hydrocephalus:   Lateral ventricles
             Subarachnoidal space enlarged
             Displacement of brain
             Ectopic heart
             Dextrocardia
             Ectopic liver
             Hemorrhage in live
             Small kidney
             Ectopic kidney
             Duodenum enlarged
             Gastroschisis

Rank 2:  Intermediate value in assessing teratogenic potential.

             Hemorrhage on olfactory bulb
             Trachea closed
             Hemorrhage in pericardium
             Hydronephrosis:  Marked
             Hydroureter

Rank 3:  Least valuable in assessing teratogenic potential
             Hydrocephalus:
             Hydrocephalus'.
             Hydrocephalus:
             Hydrocephalus:
4th ventricle
Lateral ventricles, slight
3rd ventricles, slight
4th ventricles, slight
             Nasopharyngeal canal occluded
             Nasal passage occluded
             Submaxillary gland exposed
             Trachea occluded
             Stomach distended
             Slight enlargement of kidney pelvis
             Kidney cortex solidified
             Slight enlargement of ureter
             Urinary bladder distended
             Malplaced testicle
                                13

-------
                                                   TABLE 5
                      EFFECT OF GUTHIOH EXPOSURE DURING ORGANOGEHESIS .ON THE INCIDENCE
Number of
Litters inspected
Fetuses inspected

Soft Tissue Anomalies
Hydrocephalus:  later:
Displacement of brain
Hemorrhage by olfacto-
Nasopharyngeal canal occluded
Nasal passage occluded
Trachea closed (21
Trachea occluded  (1)
Ectopic heart (3)
Dextrocardia (3)
Hemorrhage in pericardium  (2)
Ectopic liver (3)
Hemorrhage in liver  (
Stomach distended  (1)
Hydronephrosis:  mark
                 slig
Small kidney (3)
Kidney cortex solidified
Ectopic kidney (31
Hydroureter:  marked
              slight  (1)
urinary bladder distended
MaIplaced testicle  (1)
Submaxillary gland  exposed

Summary bv Rank
    1-3
  2  and  3
    3
OF SOFT TISSUE




(Rank)!/
.1 ventricles (3)
;1 ventricles, slight (1)
. ventricle (1)
. ventricle, slight (1)
ventricle, slight (1)
mlarged (3)
(3)
•y bulb (2)
iccluded (1)
I (1)




lium (2)

0

id (2)
it (1)

Led (1)

;2i
CD
ided (1)
)
jos ed ( 1 )



ANOMALIES IN RATS

0
17
109

1.2 + 1.2^
4.8 + 1.9
1.8 + 1.3
0.8 + 0.8
0 + 0
0 + 0
0 + 0
0 i 0
2.5 + 1.4
4.6 ± 1.8
0 + 0
10.4 + 4.0
0.3 + 0.3
0 + 0
0 + 0
0 + 0
1.7 T 1.7
0-0
5.9 + 4.0
3.8 + 2.1
0 + 0
0 + 0
1.7 + 1.2
1.0 ^ 1.0
0.3 - 0.8
3.3 I 2.9
9.0 + 2.9
1.0 + 1.0
48. 1+4.6
11.3 + 5.1
5.4 +2.3
Cuthion (u?/k?/dav)
1.25
19
111

1.1 +. 1.1
15.9 + 4.4-
2.1 + 2.1
6.1 + l.^
3.1 i 1.7
0^0
0 + 0
0.9 I 0.9
7.1 + 2. 3
4.7 + 2. 3
0.9 + 0.9
13.0 + 3 2
0 + 0
0.9 i 0.9
0 T 0
0^0
0 + 0
0^0
2.6 T 1.8
4.0 + 1.9
0^0
0-0
2.3 T 2.0
2.0 + 1.4
2.5 + 1.4
12.5 +4.2
4.6 ; 1.8
0^0
61.3 r 5.7
9.9 + 3.6
4.4 + 2.2
2.5
20
126

0 + 0
11.0 + 3.6
0.8 + 0.3
3.4 * 1.6
0.8 ; 0.3
0.7 + 0.7
0.7 + 0.7
0^0
6.9 * 2.3
2.5 + 1.4
' 1.0 + 1.0
11.2 + 2.8
0 *_ 0
0^0
0-0
0.3 + 0.3
3.3 - 0.3
1.7 + 1.7
4.0 + 1.6
3.0 + 1.7
0 j; 0
0+0
1.4 t 1.0
1.7 j- 1. 1
0.3 - 0.3
. 7.7 z 3.2
9.S - 3.3
0 + 0
49.7 -r 5.6
9.6 + 2.2
4.5 ; 1.6
5.0
17
92

3.9 + 3.0
15.0 + 6.6
3.9 ^ 3.0
2.0 t 1-3
1.0 - 1 . C
0 r 0
0 + 0
0 -r 0
1.3 ±1.3
2.3 t 1-5
2.9 t 2-9
17.3 + 5.9
0 t 0
0 + 0
3.9 t 3.9
0-0
0 ± 0
0^0
2.7 - 1.5
0 + 0
1.0 - 1.0
0.3-0.3
2.3 r 1.5
2.5 t. 1-
1.0 + 1.0
~. 5 ~ 1.7
6.4 - 2.7
0-0
;4 6 - 7.0
16.3 - 5.4
6.7 - 3.5
 a/   Ranked  by  increasing value in predicting teratogenic potential.
 b/   Mean *_  S.E.  of the percent of fetuses  with the indicated anomaly calculated on a per litter basis.
 c_/   Significantly different  from control (? < 0.10,  two-sample rank tast).
                                                        14

-------
oo      o i—i    CN
C;
CO
-o
oc
^j

o£
e
s- *
c
o
•r-l
1 I
pT^
O

r^
o
[ [
o
U- 1
w
PC
H

52
o

M
C/5
W
55
S3
o
o
z:
^J
^O c^
o
*ZJ
H3 o
*~2 53
23 M
S£
^
Q

W
P^
5
en
2
M
Pu

z
c
a
H

^~

P^
O

H
U
Ed
fa
fa
W
3
O










[i]
CJ

z
01
eno O cNino CN<(-
• • • •*. ••
in OOrHCNOOrHOOrHrHCnO -tfCN
CN' ^ M +1 +| +| +1 +| +| +1 +1 +1 +| +1 +| +| +| +|

ooomooooocNinrHO OCN
• • • ••• •«
CNCn rH rH i— 1 p-, inp~
rH




r*** I-H i— H c^ oo I-H CN cn
• ••• •• «.
in OOrHrHrHOrHOOOOCNrH VjCN
CN OO CN
^ ^00 +| +1 +1 +1 +1 +1 +1 +1 +1 +1 +1 +| +| +1 +|
OOinrHrHOO>OOOOr-rH CNrH
CM I-H i— i i— i vorH en rH g v-' N-X T-,
Pi ^ 3 4)
»-^ i— 1 iH -O 4J -rj
cfl -a "o cu x: c
01 M 4) l-i *^^ ^"^ *^ 00 4)
4)4)rg^-^uJrHen|j1-l 4J
1-1 4-1 3 i— 1  TJ CO rH CJ r4 "O "O Cn TJ "— v
4)4J g CJ TJ 0) 4) 4) ^s^ rq
4J4J O O 4) Q. T3 00 •• en ^,4^— ' J*:
UO C" TJ C >-i « ^ >> CN 4> C
4)4) -<034)3C4)cfli-l 4) ^ TJ 03 efl
D.O. 3 60 r- < -r4 4-1 rH 03 >, G T31-* Ed
COCO 4>rHCOO 03CO COTJl-ICflCO
CC 3Cfl05O4)-r44)r4 q-HDrHlH >
4H 1-1 1-1 o3x:cQoso'o x: •0^4-ixix: xi
O 05D.cflfflgO.iH,o3 >-
IHMCI; Ho 4)^oCC iHgt-io r,)-icO 3 cxi
Z i— 1 fa GO3nZHEV3Op3 C/3W-S3CJ c/3
03
CN i-l
CO
rH Cfl
+1
1-1
CN 4)
4J
rH 4J
i-l
rH
1-1
41
D.
rH CO

i— 1 g
A
+ 1
rH ,
rH ^£
O • Cfl C
rH g CO
+1 Cfl O M
•iH C
O 4J CO 41
C rH
4) TT O.
4-J 4) g
0 4J S
&. Cfl CO
O 1
o 1-1 o
1-1 "a S
C c -u
4) T-l
oo •
o 4) m
4J X! O
Cfl 4J •
l-i O
cu x:
4J 4J V
T-l
oo 5 cu
iH 00
4-1 4) rH
O co O
iH 3 M
TJ 4-1 4J
4) 4) C
r4 <4H O
a. o
4-4
coE
i-l O
4-1 I-l
4) C 4H
3 0)
T-H O 4J
cfl 14 c
> 4) 4)
a. i-i
00 4)
C 4) 4-1
T-I x: 4-1
03 4J iH
co *^3
0) 4-1
1-1 O rS
O rH
C • 4J
i-l Cd C
• CO
?*•* co y
XI T-l
+ 1 4-1
-a •*
4) d C
•^ cfl oo
C 4) -r-l
en n: S c/3
cd

~\ ^^. ^^
^3| Xll O|
                15

-------
                           TABLE 7
       SUMMARY OF OBSERVED SKELETAL ANOMALIES BY RANK
Rank 3:  Most valuable in assessing teratogenic potential.

             Basisphenoid displaced dorsally
             Hemi-vertebra

Rank 2:  Intermediate value in assessing teratogenic potential.

             Parietals incompletely ossified
             Squamosal split
             Squamosal incompletely ossified
             Sternebra lobed
             Split sternabrae
             Malalignment of fusion of sternabra
             Extra ossification between sternabrae
             Centri lobed
             Split centri

Rank 1:  Least valuable in assessing teratogenic potential.

             Skull collapsed slight
             Skull collapsed marked
             Nasal bones unossified
             Occipital fontanel enlarged
             Interparietal incompletely ossified
             Supraoccipital incompletely ossified
             Incus unossified
             Jugal incompletely ossified
             Hyoid bone unossified
             Hyoid bone incompletely ossified
             Unossification of sternabra
             Incomplete ossification of a sternabra
             Ribs extra
             Incomplete, ossification in pelvis
             Paws incompletely ossified
             Phalanges of paws unossified
                                16

-------
                                                IABLE  3
                  ETTSC7 OF  GUTHION  EXPOSURE  PLYING  ORGANOGENESIS ON THE INCIDENCE  OF
SKELETAL ANOMALIES IN RATS
Guthion (mg/kg/dav)

Number of
Litters inspected
Fetuses inspected
Skeletal Anomalies (Rank)-
Skull collapsed: slight (1)
narked (1)
Nasal bones unossified (1)
Occipital fontanel enlarged (1)
Parietals: incompletely ossified (2)
Interparietals: incompletely ossified (1)
Medially curved (2)
Supraoccipital: incompletely ossified (1)
3asisphenoid dorsally displaced (3)
Squaaosal: split (2)
incompletely ossified (2)
Jugal: incompletely ossified (1)
Hyoid oone: unossified 'I)
incompletely ossified (1)
Sternebrae: ossified normally
unossified (1)
incompletely ossified (1)
lobed (2>
solie (2)
aalaligned (2)
Centra: ossified normally
lobed (2)
split (2)
v'erteorae: Hemi-verteorae (3)
3.13S- excra (1)
Pelvis: incompletely ossified (1)
Paws: incompletely ossified (1)
onalanges jnossified (1)
Summarv bv lank
1-3
2 and 3
3
p_

n
117

10.3 * 5.3V
0^0
0*0
1.2 * 1.2
1.3 * 1.3
1.2 * 1.2
0^0
0^0
0 *. 0
0 r 0
2.4 * 2.4
0*0
12.9 *. 5.6
2.9 * 1.5
22.1 * 6.3
57.3 * 9.7
il. 1 * 5.5
0^0
0 r 0
0^0
90.2 : 3.5
-.1 i 2.6
3.3 ; 1.5
0*0
12.5 ^ 5."
1.2 * 1.2
5.5 ; 3.2
13.0 ; 6.2

56.3 - 5.0
13.9 * 3.9
0-0
1.25

19
123

3.3 - 3.3
1.4 i 1.3
0 + 0
:.5 i 1.9
0.7 * 0.7
2.7 i 1.5
0^0
0.3 i 0.3
0^0
0^0
3 - ^
0 ; 0
7.9 t 3.9
0.7 r 0.7
25.3 - 5.7
53.9 ^ 3.1
36.9 - 4.1
1.3 * 1.3
1.3 z 1.3
1.6 — 1.1
77.3 ^ 5.1
13.9 - 5.3
5.7 - 1.3
3.7 r 0.7
3.3 - 1.5
1.5 - 1.1
5.2 - 4.5
16.0 -7.3

34.5 ±4.5
23.5 - 5.0
0.7 - 0.7
2.5

20
136

3.1 i 1.3
1.7 * 1.7
0*0
0*0
2.3 * 1.3
1.0 ^ 1.0
1.4 * 1.4
0*0
0 ; 0
3-0
0*3
^ — '^
13.9 - & . 3
0.7 - 0.7
27.6 t 6.3
48.1 - 7.3
46.6 z 4-9
0.7 - 0.7
0-3
T 2 — " 2
30.0 * 5.5
16.5 - 4.5
4.9 ; 2.1
3 -; °
4.1 - 1.7
1.0 - 1.0
3.0 - 3.0
7.5 — 3.5

T.2 - 5.3
25.2 - 5.1
3-0
5.0

17
101

11.9 - 6.3
0 Z o
1.0 j; 1.0
2.0 - 2.0
0.3 - 0.3
1.3 * 1.0
0*0
0-0
3.6 - 2.3
1.6 - 1.1
0^0
0.7 ^ 0-7
3.3 - 3.5
1.7 ~ 1.7
20.5 ; 5.7
*2.7 ^ 7.4
56.9 * 6.2
3-3
1 ; 0
"-»'"'
90. 3 - 3.6
5.4 - 2.4
1.7 - 1.2
•^ 1
J — 'J
9.9 - 2.9
2.9 - 3.9
2.9 - 2.9
9.9 ; 6.1

3ft.- - -.2
1" . 3 - 4.. 3
3 - 0
a/  Ranked by increasing value  in oredicting  :eratogenic  potential.
by  Mean * S.E. of the percent  of fetuses with  tr.e  indicated  anomaly calculated ;t
per litter basis.
                                                         17

-------
                           o

                           m
ON

w
rJ
         w
         a
         Q
         r-4
         CJ
         £9
         H
         z   w
         O   CJ
Cd

W   00
O   Cd
O
Z
Pi   O
o   -z

o
z
M
         Q

         Pd


         CO
         z
         o
         M


         1
         H
         CJ
         Cd

         fc
         W
                       CO
                      •U
                       oe
                       c
                       o
                       o
                                        t*^  m
                                        r-l  OO
                               oo  en
                               r-4  O1
                           0|
                                             oo
                                     "4-J
                                     O


                                     CU


                                     E
                                          y   o
                                          4)   4)
                                          CO   CO
                                          C   C
                                 CO   CO

                                 4)   CO






























































cSl
,*-"V.
££
a
CO
&*
^— '

CO
4)
i-4
i—l
CO
E
C
C


^
.jj
cu
i— i
OJ
_^
en

^
en
4-1
»
^j-


m
•
CM

4-i
m
en

o
4-i
y3
•
en




o
•
i—4
4-1

*cj-
•
en



























/^v
i— i
^^

4-1
r!
ciO
•H
r^
CO


..
TJ
CU
CO
d.
CO

i— i
O

1— 1
I— <
3

en


0
4-1
o





o

4-1
0


r-i
4-1
,_<
•
r-4






o
4-1

O
















s-, .
r-l
swl

TJ
CU
i-l
M-l
i-l
co
co
O

^^
r-4
4)
4J
CU
i— 1
Ccu
E
Q
y
C
•rl


* .
1— 1
ed
4-1
4)
•i-l
V4
Q.
^i
4)
4-1
rj
M

-^
r-4
4-1
•*
r-l




O

4-1
0



4-1
o








o
4-1

o






















s-\
CN
**-•

T3
0)
^>
^4
3


r^»
f— 1
T— 1
cj
•*•(
T3
OJ
2
















^
on o o
+ 1 4-1 -f 1
CO 0 0
vj-


\o o
• •

1— i
CU <~v
-LJ (-H
^
rH
CL *T3
0 /— x QJ
O^*^
o ^^^ ^-*
C *"
•-4 T3 CO
4) co
1-4 O
. . 14_| £2
r-4 -rl 3
03 CO
4J CO
1-4 O ••

i-l 3 C
y o
o ••
ca co "o
V4 3 "rl
a. y o
3 C ?^
en '-' —

in
m
4-1
oo
m
"

r-H
*
^O

4-1
00
0

r~-
4-1
yO
.
>
— t
(™H
cd


o
C

"O
-( <)• O O 
O m o tn •— 4 o CA ^^
. . . . O
^ ^— •* *H r— *
r— i M-! r— 1
T3 3J "O T-| Gj

3
si
CO
1 -i-4
CO
3 CO
_n
rl ^
f CU
• 4-1
-1 4-1
4)
(X
!•' C
hi °
2 4)
• 4J
i ca
3
y --^
cd co
y 4)
*4 Jx,
. . i— I ^
"t r-l CO C
, ca E CO
hi 1-1 O l-i
4J C
t C CO 4)
4) r-l
3 4J T-J O.
O 4) E
a. 4-1 ca
CO co
y y i
i-l i-l O
c -a 3
4) C 4J
00 i-l
o
4J 4) m
eo x: o
4) O
4J .2
4-1 V
00 —4
G S C— 4
-^ j ^ _*
4-" CO
y 4) 1-4
•rl CO O
T3 3 rl
4) 4J 4-1
1-4 4) C
a u-4 o
y
C HH
1-1 o E
O
4) 4-1 (-1
3 C H-i
r- CU
co y 4-1
> 1-4 C
4) CJ
so a, ^
C 4)
i-l 4) <4-4
CO jC M-4
C8 4-1 --I
4) T3
(-1 M-l
y o >,
C r-l
1-1 . 4-1
M C
>, . TJ
Xi cn y
—i
"O 4-1 ^-i
4) -4
I ^ C C
•3 CJ — 1
si S cn


•^ -^^ -. 	
c| x:l y
                                                                               13

-------
                               TABLE 10
     EFFECT OF GUTHION ADMINISTERED DURING GESTATION AND LACTATION
ON MATERNAL WELFARE AND REPRODUCTION


Rats Treated
Non-pregnant (%)
Dead (%)
Pregnant (%)
Dead - total (%)
Days 6-16
Day 16 - delivery
Postpartum
Feed Intake (gm/rat/day)
Gestational days 6-20
Postpartum days 7-21
Body Weight (gin/rat)
Gestational
Day 0
Day 6
Day 16
Day 20
Postpartum
Day 0
Day 4
Day 7
Day 14
Day 21
Duration of Gestation

0
14
1(7)
0(0)
13(93)
0(0)
0
0
0

27 + 1
68 + 4


241 + 4
275 + 4
331 + 7
387 + 9

311 + 9
304 + 8
317 + 8
346 + 8
342 + 10
21.6 +0.1
Guthion
1.25
14
1(7)
0(0)
13(93)
0(0)
0
0
0

26 + 1
64 + 5


241 + 4
276 + 5
333 + 7
382 + 11

297 + 7
304 + 7
316 + 7
344 + 7
341 + 6
21.8 + 0.1
IN RATS
(mg/kg/day)
2.5
14
2(14)
0(0)
12(86)
0(0)
0
0
0

28 + 1
63 + 6


248 + 4
286 + 5
345 + 4
400 + 10

315 + 6
320 + 5
332 + 4
363 + 4
360 + 5
21.9 + 0.1

5.0
15
2(13)
2(100)
13(87)
8(62)
3
4
1

20 + IL-
47


239 + 4
276 + 5
293 + 8^.'
327 + 13^

260 + ll-''
257 + 14-/
278 + 21
340
346
22.0 + 0.2
(days)
a/  Significantly different from control (Dunnett's procedure).
                                     19

-------
                               TABLE 11

             EFFECT OF GUTHION ADMINISTERED TO DAMS DURING
GESTATION AND LACTATION
ON PUP
WELFARE
Guthion (mg/kg/day)

a/
Viable Litters-
Birth
Day 4
Day 7
Day 14
Day 21
Total Implants /Dam-
Pups Delivered/Dam
Live Pups /Dam
Birth
Day 4
Day 7
Day 14
Day 21
Pup Weight (gm/pup)
Birth
Day 4
Day 7
Day 14
Day 21








12.
11.

11.
11.
11.
11.
11.

7.
9.
12.
24.
37.
0


13
13
13
13
13
5 + 1.
8 + 1.

7 + 1.
6 + 1.
2 + 1.
0 + 0.
0 + 0.

1 + 0.
0 + 0.
0 + 0.
5 + 1.
0 + 1.








1
1

1
0
0
9
9

5
4
7
1
6








13.
11.

11.
10.
10.
9.
9.

6.
8.
12.
23.
37.
1.25


13
13
13
13
13
8 + 0.
9 + 0.

8 + 0.
2 + 1.
0 + 1.
9 + 1.
9 + 1.

4 + 0.
8 + 0.
5 + 0.
7 + 1.
0 + 1.








6
8

8
3
3
3
3

3
6
8
2
8








13
12

12
11
11
11
11

6
8
12
22
34
2.5


12
11
11
11
11
.9 +
.3 +

.2 +
.6 +
.5 +
.4 +
.4 +

.5 +
.6 +
.2 +
.8 +
.4 +








1.
1.

1.
1.
1,
1.
1.

0.
0.
0.
1.
1.








1
3

2
5
5
4
4

2
4
6
1
6
5.0


6
3
1
1
1
13.4 + 0.9
11.2 + 0.8

10.7 + 1.1
11.7 + 0.3
10.0
5.0
5.0

5.7 + 0.2^
5.4 + 0.5^'
7.8
14.4
24.4
a/  Number of litters with at least one viable pup.
b_/  Determined by uterine scarring on postpartum day 21.
c/  Significantly different from control (two-sample rank test) .
                                   20

-------
                               TABLE 12
           REPRODUCTIVE INDEXES OF FEMALE RATS GIVEN GUTHION
DURING GESTATION AND LACTATION
Guthion (mg/kg/day)

Fertility-7
Gestation—
c/
Pup Survival-
Day 0-4
Day 0-7
Day 7-14
Day 4-21
0
93 (66-100)
100 (75-100)

100 + 0
97 + 1
98 + 1
96 + 2
1.25
93 (66-100)
100 (75-100)

86 + 9
85 + 9
100 + 1
'95 + 4
2.5
86 (57-98)
100 (73-100)

87 + 9
86 + 9
99 + 1
98 + 1
5.0
87 (59-98)
46 (19-75)

46 + 26^
14 + 14^
50
14 + l47
a/  Confirmed pregnancies/sperm-positive females x 100  (95% confidence limits)
b_/  Confirmed pregnancies with viable pups/confirmed pregnancies x  100 (95%
      confidence limits).
cj  Percent of pups surviving during the indicated interval.
d/  Significantly different from control (two-sample rank test).
                                      21

-------
                                  TECHNICAL REPORT DATA
                               ' read Instructions on rlie reverse' before completing/
1  REPORT NO
EPA-600/1-78-056
4. TITLE AND SUBTITLE
 TERATOLOGY OF GUTHION
                                                          3 RECIPIENT'S ACCESSION>NO
                                                          5, REPORT DATE

                                                             August 1Q78
                                                          6. PERFORMING ORGANIZATION CODE
7 AUTHOR(S)
 Robert  D.  Short,  Jan L. Minor, Timothy  M.  Unger, and
 Cheng-Chun Lee
                                                          8. PERFORMING ORGANIZATION REPORT NO.
9. PERFORMING ORGANIZATION NAME AND ADDRESS
 Midwest Research Institute
 425  Volker Blvd.
 Kansas  City,  MO 64110
             10. PROGRAM ELEMENT NO.
                1EA615
             11. CONTRACT/GRANT NO.
                68-02-2746
12. SPONSORING AGENCY NAME AND ADDRESS
Health Effects Research Laboratory
U.S.  Environmental Protection Agency
Office of Research and Development
Research Triangle Park, N.C. 27711
                                                          13. TYPE OF REPORT AND PERIOD COVERED
   RTP,NC
             14. SPONSORING AGENCY CODE
                EPA 600/11
15. SUPPLEMENTARY NOTES
16. ABSTRACT
      The purpose of this study was  to  assess the effects of Guthion,  a pesticide
with anticholinesterase activity, on development in rats and mice.   A preliminary
toxicity study with Guthion indicated  that a 35 LD5_ dose for virgin rats and a 10 day
LD5Q dose for virgin mice was between  4  and 8 mg/kg/day for both  species.  On the
basis of this data, doses of 0, 1.25,  2.5, and 5.0 mg/kg/day were selected for the
developmental study, which consisted of  two phases.  During the first phase, pregnant
rats and mice were treated for 10 days starting on gestational day 6.   The high dose
affected maternal welfare only in rats.   Guthion did not significantly increase in a
dose-related manner any of the specific  anomalies observed in either rats or mice.
During the second phase, pregnant rats were treated from gestational day 6 to post-
partum day 21.  Dams in the high dose  group were more sensitive to Guthion later in
gestation with the result that deaths  and signs of anticholimjsterase toxicity
increased during this time.  Guthion also adversely affected maternal welfare in this
group.  As a result of Guthion toxicity,  only one litter survived until weaning. The
inability to dissociate toxicity in adult and developing animals  suggests that Guthion
has little primary effect on the development of rats or mice.
17.
                               KEY WORDS AND DOCUMENT ANALYSIS
                 DESCRIPTORS
b.IDENTIFIERS/OPEN ENDED TERMS  C. COSATI Field/Group
  pesticides
  cholinesterase inhibitors
  Guthion
  teratology
 06 T
   •>ISTRIBUT,O\ STATEMENT

    .EASE TO PUBLIC
19 SECURITv CLASS I This Report)
  UNCLASSIFIED
21 NO. OF PAGES

  25
                                             20 SECURITY CLASS /This page;
                                               UNCLASSIFIED
                                                                        22 PRICE
      l 2220-1 (9-73)
                                            22

-------