EPA-600/1-78-004
January 1978
Environmental Health Effects Research Series
EVALUATION OF NEONATE SQUIRREL MONKEYS
RECEIVING TRITIATED WATER
THROUGHOUT GESTATION
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
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.
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EPA-600/1-78-004
January 1978
EVALUATION OF NEONATE SQUIRREL MONKEYS
RECEIVING TRITIATED WATER
THROUGHOUT GESTATION
by
David C.L. Jones
Stanford Research Institute
Menlo Park, California 94025
Contract No. 68-02-2280
Project Officer
James F. Wright
Experimental Biology 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.
This particular project was conducted to assess the effects of
continuous low-level intrauterine exposure of subhuman primates to
an energy-related pollutant.
John H. Knelson, M.D.
Director,
Health Effects Research Laboratory
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PREFACE
The diversity of disciplines and efforts required for this project
are reflected by the scientific and technical personnel who contributed
their talents and dedication. The procurement and maintenance of the
animals were directed by Samuel J. Jackson, D.V.M., who was assisted by
his Animal Care Services Staff, particularly Richard V. Romero,
Arthel Coleman, and Clarence Rhodes. Evaluation of reproductive cycle,
breeding, and pregnancy testing were directed by Roger A. Earth, D.V.M.,
and Sidney J. Stolzenberg, Ph.D., with assistance from Shirley M. Madan,
Deborah K. Palmer, and Victoria E. Gorum. John S. Krebs, Ph.D., supervised
the preparation and administration of the tritiated water. Chozo Mitoma,
Ph.D., directed the assays of the tritiated water and of the animal
tissues and urine for radioactivity; he was assisted by Thomas Steeger
and Jeanne Kalivoda. Daniel P. Sasmore, D.V.M., assisted by Sandra J.
Phillips and Barbara A. Kirkhart, was responsible for necropsy, histology,
and diagnosis of tissues. Joel N. Kaplan, Ph.D., generously advised on
animal maintenance and breeding strategies. In addition to the SRI staff .
participants, R. Lowry Dobson, M.D., of the U.C. Lawrence Livermore
Laboratory contributed invaluable advice on the interpretation of the
histology of the ovary.
iv
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CONTENTS
LIST OF TABLES vi
INTRODUCTION 1
SUMMARY AND CONCLUSIONS 3
RECOMMENDATIONS 5
PROCEDURES AND RESULTS 7
Animals and Breeding Procedures 7
Administration of HTO 10
Evaluation of Pregnancy 13
Body Weight 14
Births 16
Concentrations of HTO in Body Water 18
Assay of Radioactivity in Tissues 23
Gross and Microscopic Evaluations of Progeny 24
Hematology and Serology 30
REFERENCES 59
TECHNICAL REPORT DATA FORM 61
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TABLES
1 Scheduled HTO Values 33
2 Mean Assayed HTO Drinking Water Activity 34
3 Pregnancy Evaluation Data for Inseminated Female
Squirrel Monkeys Receiving HTO In Drinking Water 35
4 Body Weights of Inseminated Female Squirrel Monkeys
Not Providing Full-Term Deliveries or Evidence
of Frank Abortion 36
5 Body Weights of Female Squirrel Monkeys Delivering
Full-Term Progeny 37
6 Time Distribution of Full-Term Births 38
7 Estimates of Gestation Period for the Squirrel Monkey . . 39
8 Mean HTO Concentrations in Urine of Inseminated Adult
Female Squirrel Monkeys 40
9 Concentrations of HTO in Urine of Control Animals Not
Receiving HTO in Drinking Water 41
10 Urinary Concentration of HTO in Group 1 Adult Squirrel
Monkeys Delivering Full-Term Progeny 42
11 Urinary Concentration of HTO in Grpup 2 Adult Squirrel
Monkeys Delivering Full-Term Progeny 43-
12 Urinary Concentration of HTO in Group 3 Adult Squirrel
Monkeys Delivering Full-Term Progeny 44
13 Urinary Concentration of HTO in Group 4 Adult Squirrel
Monkeys Delivering Full-Term Progeny 45
14 Urinary Concentration of HTO in Group 5 Adult Squirrel
Monkeys Delivering Full-Term Progeny 46
15 Urinary Concentration of HTO in Group 6 Adult Squirrel
Monkeys Delivering Full-Term Progeny 47
16 Urinary Concentration of HTO in Group 7 Adult Squirrel
Monkeys Delivering Full-Term Progeny 48
17 Analysis of Mean HTO Concentration in Urine of Pregnant
and Nonpregnant Female Squirrel Monkeys 49
18 Tissue Radioactivity of Neonate Squirrel Monkeys 50
19 Tissue Radioactivity of Neonate Squirrel Monkeys
After Dehydration 51
vi
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20 Linear Regression Analysis of Tissue Radioactivity
in the Neonate Squirrel Monkey .............. 52
21 Distribution of Histologic Lesions Among Full-Term
Neonate Squirrel Monkeys ................. 53
22 Body Dimensions and Organ Weights in Neonate Squirrel
Monkeys by HTO Concentration ............... 54
23 Body Dimensions and Organ Weights of Male Neonate
Squirrel Monkeys ..................... 55
24 Body Dimensions and Organ Weights of Female Neonate
Squirrel Monkeys ..................... 56
25 Hematologic Parameters of Neonate Squirrel Monkeys .... 57
26 Serologic Parameters of Neonate Squirrel Monkeys ..... 58
vii
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INTRODUCTION
The objective of this project was to provide information that would
be pertinent to evaluating the hazards to the general population of
tritiated water (HTO) in the drinking water supplies. This situation
could arise during the reprocessing of nuclear fuel and the mixing of
processing waste streams with rivers providing drinking water to cities
and, to a lesser extent, from the production of HTO in nuclear reactors
and release to the environment.
Because previous studies with rats had shown that delayed reflex
development and hypoactivity were induced by exposure to low levels of
radiation from conception throughout life, the squirrel monkey (Saimiri
sciureus) was selected as the test animal in which to evaluate the
potential HTO hazard in primates using, primarily, behavioral indices of
effect. Drinking water levels were to range from about 16 to 1300 times
the accepted permissible concentration (0.003 yCi/ml) in drinking water
for the general population; these concentrations were to be administered
to pregnant female squirrel monkeys from conception to either birth or
weaning of their progeny.
After the project had proceeded into the stage of administering HTO
to inseminated monkeys, the primary focus of assessing the effects was
changed from evaluating alterations in behavioral development of the prog-
eny during the first postnatal year to conducting a physical and histologic
evaluation of the progeny at birth. Criteria included, gross and micro-
scopic pathologic effects, body conformation and organ weights, and tissue
distribution of radioactivity in the progeny and reproductive parameters
in the. inseminated adults. Still later certain ancillary studies were
incorporated that involved specialized brain histology procedures and
evaluation of ovarian changes in the progeny by estimating the total
primary oocyte count. The latter studies, being conducted at other labora-
tories, have not yet been completed, so only preliminary results for these
evaluations can be presented at this time. The primary studies, conducted
entirely at SRI, have been completed and form the basis for this final
report.
1
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SUMMARY AND CONCLUSIONS
The proportion of squirrel monkeys bred under the conditions of this
study expected to deliver full-term progeny was overestimated by a factor
of two to three. Both the proportion of inseminated animals that actually
conceived and the proportion of pregnant animals that provided full-term
progeny were lower than expected. In addition, the ratio of the HTO
concentration in body water to that in drinking water was lower than
expected for the squirrel monkey, so that body water concentrations were
about 75% of the initially scheduled levels. Despite these differences
from the initial design-parameters, several conclusions about the
effects of HTO administration during gestation appear to be valid.
The present data indicate no discernible anatomic or physiologic
effects on the neonate squirrel monkey of HTO administeration to the
mother throughout gestation in drinking water at levels ranging from
16 to 1,000 times the permissible level for consumption (0.003 uCi/ml).
The neonate monkeys were unaffected in terms of the following criteria:
Body weight
Selected body dimensions
Selected organ weights
Hematologic patterns
Histology of selected organs and tissues other than ovaries.
The number of primary oocytes in female neonate squirrel monkeys apparently
decreased markedly with increasing levels of HTO in the drinking water
during gestation. The need for quantifying this effort was not foreseen
in planning this project, and the appropriate technique is laborious and
exacting. Nonetheless, neonate ovaries are being processed and evaluated
by a collaborator (Dr. R. L. Dobson, U.C. Lawrence Livermore Laboratory),
who expects the results will be available within 3 months.
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Based on the following criteria, HTO administered at the levels
specified either throughout pregnancy or, in the case of animals not
delivering full-term progeny, for the equivalent time (about 22 weeks)
had no discernible effect on inseminated adult squirrel monkeys:
Abortion and/or resorption rate
Gestation period
Body weight.
The effect of HTO administration on the number of primary oocytes
in the adult female squirrel monkey is not known. Unilateral ovariectomies
were performed on a total of 56 females from the controls and the four
highest dose groups; these tissues are now being processed for oocyte
counting by Dr. Dobson. Because the time requirements for this effort
are even greater than those for evaluating neonate ovaries and because
present resources are limited, these results will not be available until
about a year from now.
Certain information not pertaining to the effects of HTO administra-
tion was also generated from this project. Timed-pregnant squirrel
monkeys can be provided from a nonhabituated animal population wherein the
social structure of the breeding groups is not strongly stabilized;
however, the full-term delivery rate may be reduced as compared with
more conventional breeding practices. Identification of inseminated
animals using the presence of a vaginal plug as the criterion and identi-
fication of pregnant animals using the mouse uterus weight procedure
yield essentially no false negatives and the former is a much less time-
consuming procedure than vaginal slide preparation and examination.
The insemination rate falls markedly at the beginning of the second
week of continuous breeding and becomes very low after the second week.
With the present procedures, both the pregnancy rate of those inseminated
and the resorption rate of those pregnant were of the order of 50%.
For squirrel monkeys of Bolivian origin, the gestation period appears
to be between 141 and 159 days, with a mean of 152.5 days (S.D., 3.9 days).
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RECOMMENDATIONS
1. Upon completion of the evaluation of the quantitative data on oocyte
counts in the neonate squirrel monkeys, the pertinence of this
information to the potential hazard to human population of HTO in
drinking water should be assessed.
2. Similar considerations should be made after the quantitative oocyte
data for the adult squirrel monkeys have been evaluated.
3. The need for further studies involving HTO administration to primates
to resolve questions arising in the above evaluations should be
considered.
4. A means for following the future reproductive performance of the
adult females exposed to HTO at SRI and now being held at the
Caribbean Primate Research Center should be implemented as soon
as possible.
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PROCEDURES AND RESULTS
Animals and Breeding Procedures
Although most squirrel monkeys (Saimiri sciureus) used in research
in the United States come from Columbia or Peru, Bolivia was the only
source that had a sufficient number of adult female monkeys for this
project at the time of its initiation. Accordingly, arrangements were
made with a commercial importing company to provide a total of 400 adult
females from that country. Only females with external indications (such
as nipple development) of having previously delivered offspring were
accepted. This large number of females was considered necessary because
the objective was to provide at least 100 viable 6-month-old progeny,
and loss rates due to failure to become inseminated, failure to maintain
pregnancy, and/or to infant mortality can be high, particularly in
unacclimated, newly imported animals. In addition to the 400 females
(ten cohorts of about 20 to 50 each), 40 adult males were also acquired
from the same source.
After the usual 5-week quarantine, the females were assigned to
groups of 15, and each group occupied a concrete-floored wire-mesh run
(1.2 m x 2.4 m * 2.3 m). The males were housed similarly, but separately,
before breeding. For this report, breeding means only that males and
females were accessible to each other. Tap water was supplied ad libitum
via automatic drinking valves. Vitamin supplement"'" was provided in
1-liter drinking bottles twice a week. Feed^ was supplied twice a day.
The floors of the pen were hosed daily with tap water.
Primate Imports, New York.
Tang , General Foods Corp., and Visorbin3', Norden Laboratories.
Monkey Chow 25, Ralston Purina Co.
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The squirrel monkey has a 5- to 14-day reproduction cycle that con-
tinues for about 3 months; the particular time of year of the cycle varies
according to geographic latitude and length of time of residence at a par-
ticular latitude. Once some females in a given colony begin their cycle,
the proportion of the females in cycle increases rapidly. The method for
evaluating the reproductive state depends on the presence of nucleated
epithelial cells and/or cornified cells in vaginal washings, these cells
being indicators of the occurrence of cycling. A pipette containing
physiologic saline is inserted into the vagina, the solution is rinsed
in and out, and an aliquot is transferred to a microcell for reading at
400xmagnification. In the case of breeding animals, the presence of
sperm in the slide indicates insemination. Some females of the .first
cohort were assessed daily for cycling as soon as they had completed
quarantine in late April. Some of them were found to be in cycle, and,
by the time that a sufficient number of animals had become available to
begin breeding (late May), all the animals were in cycle.
During the cycling evaluation, which continued for about a month
as a means of evaluating for insemination, we discovered that over 90%
of the females showing sperm in the vaginal washings also had a seminal
coagulum or plug in the vagina, detectable visually or by observing the
flow characteristics of the wash fluid during vaginal rinsing. Because
detection of plugs was much more efficient in terms of technician time,
insemination was defined as the presence of a plug, and the preparation
of vaginal slides was discontinued.
An important objective of the breeding effort was to identify
inseminated monkeys within 24 hours of insemination so that HTO adminis-
tration could commence essentially coincident with conception. For
economy of operations, accurate recognition of insemination was important.
Inaccuracy could have resulted in beginning HTO administration to
uninseminated monkeys or in identifying inseminated animals at a later
date, thus delaying HTO reception until significantly later in gestation.
In the squirrel monkey, subsequent insemination can occur many days after
conception. Because of this explicit need to initiate HTO administration
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at conception, personnel had to be available to detect insemination in
every breeding female within 24 hours of possible insemination. Because
the same personnel were also conducting the pregnancy evaluations in
already inseminated animals, and because we realized that ultimate
evaluation of progeny would be more efficient with a consistent birth
schedule as opposed to a sudden wave of births, the breeding schedule was
adjusted accordingly.
Initially, some females were placed in individual cages one-to-one
with a male, but this method appeared to yield no more inseminations than
did a breeding unit consisting of 15 females and 3 males in a pen. As
the project progressed, inseminated females were removed for HTO adminis-
tration and either naive females were added to the breeding pens or groups
were combined to maintain an efficient group size. As pregnancy testing
became a major effort, breeding was interrupted whenever personnel were
unavailable to check for insemination within 24 hours. Thus, the
precise breeding history varied among animals.
Of the 406 females received, 373 were bred. The remaining 33 were
found dead before breeding (N = 10), were immature (N = 14), were preg-
nant upon arrival (N = 2), were injured during their scheduled breeding
period (N = 3), or were not permitted to breed through oversight (N = 4).
Of the 373 animals bred, 277 (74%) were inseminated. Of these, 242 were
inseminated as a result of continuous breeding, and the remaining 35 were
inseminated through intermittent breeding. The 96 noninseminated animals
were bred for a median of 23 days (range, 2 to 46 days) without detectable
insemination. None of these animals either delivered or aborted within
the possible gestation period since the last breeding day. For the animals
inseminated during.continuous breeding, the median time to achieve
insemination was 5 days, with 25% of the monkeys inseminated within
2 days and 75% within 8 days. Sporadic peaks occurred in the daily
insemination rate (proportion of the animals still in the breeding situ-
ation that were inseminated) during the next 10 days, and one animal was
inseminated after 37 continuous days of breeding. However, the probability
of insemination clearly became much lower after about 2 weeks.
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Administration of HTO
Three shipments of high specific activity HTO were acquired from two
suppliers: in May* 1 Ci and 7 Ci in May and 3 Ci in October.f Stock solu-
tions were prepared by successive dilution, and appropriate aliquots of the
stock were transferred to capped vials for final dilution with tap water.
Aliquots of the stock solutions were diluted with physiological saline in
sterile serum bottles for initial injections into inseminated monkeys.
The objective was to bring the concentration of HTO in the body water
rapidly up to the desired level by means of intraperitoneal (ip) injec-
tion on the day of insemination and then maintain it throughout gestation
by administration in the drinking water.
The desired body water HTO concentrations were 0 (control), 0.05,
0.2, 0.5, 1.0, 2.0, and 4.0 yCi/ml for the seven groups. Calculation of
the microcuries to be injected initially was based on the average weight
of a sample of the animals at that time (660 g) and on an estimated mean
total body water content of 455 ml (weight x 0.69). Initial scheduled
concentrations for administration of HTO in the drinking water were based
on the estimate that the body water HTO concentration would approximate
83% of the drinking water concentration. Radioassay during the first few
weeks of administration indicated that the actual HTO concentrations in
the drinking water were lower than expected, apparently because the
initial 1-Ci shipment was mislabeled. Accordingly, scheduled drinking
water concentrations were increased after the fourth week. During the
next 2 weeks, by which time sufficient urine samples were available for
a more precise evaluation, the initial estimate of the urine-to-drinking
water ratio of 83% was found to be too high, even with large variabilities
in urine radioactivity both within and between groups. Thus, the
scheduled HTO concentration in drinking water was increased again at the
end of the sixth week.
New England Nuclear Co.
Amersham-Searle Co.
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The lower-than-expected'concentrations of HTO in body water appeared
to be related primarily to a combination of maintenance procedures and
squirrel monkey behavior. After food pellets were put into the hopper,
the monkeys began eating, holding a pellet in the forepaws and frequently
dropping it on the floor. About an hour later, the animal technician
sprayed the floors of the pens to presoak the accumulated food debris and
excreta and then returned later to hose the floors clean. In the interim
between soaking and hosing (about a half-hour), the monkeys had access to
tap-water-soaked pellet debris. Thus, total water intake included a vari-
able fraction of nontritiated water. This probably accounts for the
initial underestimate of the body water-to-drinking water ratio and also
probably contributed significantly to the observed variability within and
between animals. Table 1 summarizes the pertinent data.
On the day that insemination was detected, females were assigned to
one of the seven groups (by use of a table of random numbers), received
1 ml of the appropriate HTO solution, and were transferred to a pen in
another building where the appropriate concentration of HTO in the drink-
ing water was available. Animal maintenance procedures and the pen
dimensions were as described above. Tritiated drinking water was prepared
initially by rinsing one of the stock solution vials into a 25-liter
polyethylene carboy, diluting with tap water, and mixing by rotation.
Water from the carboy was withdrawn into 1-liter glass bottles with
conventional stainless-steel drinking stems, which were then fastened to
the cyclone-wire pen doors. Controls received tap water in similar
bottles. As the number of inseminated monkeys increased, about 60 bottles
were required per week. Because of the many hours spent handling bottles
and because of the associated potentially large error possibilities, the
HTO administration system was converted to 20-liter tanks fastened
directly to the pen doors, with a drinking valve on the inside of the
cage. Controls drank tap water from a drinking valve attached directly
to the tap water supply pipe. Each of the 20-liter tanks was calibrated
and marked at 5-liter intervals, and the stock HTO vials were rinsed
directly into the tanks and diluted with a high-pressure hose, fitting so
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that mixing was accomplished through churning. Twice weekly, vitamin
supplements were supplied in 1-liter bottles by dissolving the supplement
in HTO drawn from the appropriate tank.
The radioactivity of drinking water was determined on duplicate
aliquots of 0.1 ml in a liquid scintillation system. Quench compensa-
tion was made by automatically obtaining external standard pulse height
(ESP values) for each sample. The ESP values were used to correct the
count of the colored sample to that of the unquenched sample by reference
to a standard curve obtained by plotting the degree of quench as a function
of ESP values. All samples were counted in polyethylene vials. The
tritiated water standard was counted each time to calculate the counting
efficiency. The efficiency varied between 46.5 and 56.8%, with a mean
of 50.8 and a standard deviation of 2.3%. The variation in the counting
efficiency is partly a function of the scintillation fluid used.
Table 2 presents the results. The missing entries for the weeks
of 7/9, 7/16, 8/13-8/27, and 9/3 were associated with changes in the
operation schedule. The sample from Group 2 for 10/8 was not assayed
by error. Only animals in Groups 2 and 3 were still receiving HTO during
the last week. The changes in scheduled HTO concentration noted above
are reflected in the lower values during the first 6 weeks. Beginning
with the week of 7/23, the mean HTO concentrations were within 5% of
the scheduled values (Table 1), and almost all values were within one
standard deviation of the mean. Standard deviations increased with
increasing HTO concentration and ranged from 4 to 13% of the mean.
Searle Analytic, Inc., Mark III.
Packard Instrument Company, Inc.
f TM
New England Nuclear's Oxifluor-H20 was used first, but later Aquasol
was used because it gave a slightly higher efficiency.
10
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Evaluation of Pregnancy
Evaluation for pregnancy in inseminated monkeys comprised three
objectives. The first was to provide a means for selection in case
replacing an inseminated but nonpregnant animal with another subsequently
inseminated monkey was necessary because of space limitations in the pens
reserved for HTO administration. Second, pregnancy evaluation was to
provide data on the rate of resorption* as a function of HTO dose level,
if such a relationship existed. Third, pregnancy evaluation was to
provide a check on the adequacy of insemination, as defined above, and
on the breeding regimen.
The bioassay method used for pregnancy determination was essentially
similar to the methods customarily used,3''* but it was slightly modified
so that large numbers of assays could be performed. An aliquot of each
of the 24-hour urine samples collected for HTO assay from the inseminated
monkeys was used for pregnancy assay. Three 10- to 12-g female mice were
injected ip once daily for 3 days with 0,3 ml of the urine. One control
group of mice received 0.3 ml of physiological saline injected ip, and
a second control group of mice received no treatment for each set of
urine samples tested. On the fourth day, all mice were killed by
cervical dislocation, and the uteri were removed, stripped of fat and
connective tissue, pressed on a paper towel to remove luminal fluid, and
weighed to the nearest 0.2 mg on a torsion balance.^
To avoid discarding pregnant monkeys from the study, we initially
considered that a 33% increase in the uterine weight of urine-treated
mice over that of control mice indicated pregnancy. This was
based on criteria used by Wilson and Fradkin* who injected monkey serum
into five mice. Ultimately, we found that a 100% increase in uterine
weight of urine-treated over control mice was a better index of pregnancy
in our study and used this as the criterion of a positive pregnancy test.
Resorption is used in this report to distinguish an animal that was
positive on the pregnancy test but failed to provide either frank evi-
dence of abortion (fetal, or placental material, vaginal bleeding) or
a full-term delivery.
TRoller-Smith.
11
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In some cases, no reliable result was obtained either because of
insufficient urine or because the urine was toxic to the mice, presumably
because of contamination by food and/or feces in the collection system,
although even urine that appeared clean was occasionally toxic. Weekly
samples were available for most animals, however, and Table 3 shows the
data for all animals evaluated. These data indicate that about half of
the inseminated monkeys had conceived and that resorption or frank abor-
tion occurred in over half of the pregnant monkeys. The major portion
(70 of 78) of the total abortions were resorptions. Because pregnancy
testing generally did not continue past the sixth week postinsemination,
we cannot specify when resorptions tended to occur for these 70 animals.
For the 10 animals in which gross signs were apparent, the abortions
occurred between 78 and 129 days postinsemination.
Body Weight
Inseminated animals were weighed at approximately 4-week intervals
during the period of HTO administration. Because of the introduction of
animals to each dose group as they became inseminated, the number of
animals weighed during any particular week varied. For most of the experi-
ment, two dose groups were weighed each week (one on Monday or Tuesday,
the other on Wednesday or Thursday) just before being placed in metabolic
cages for urine collection. Because seven groups were used, only the
controls were weighed during the fourth week of the weighing cycle. As
the experiment progressed, the precise weighing schedule was varied some-
what according to staff availability, so that in some cases some animals
were weighed twice within 4 weeks and were not weighed during the next
4-week interval.
Body weight obviously is affected by pregnancy, particularly after
the first trimester. Table 4 presents the data for animals that did not
provide full-term deliveries or evidence of frank abortion and Table 5
presents the data for those that did produce full-term progeny. These
data in Table 4 indicate that body weights in the squirrel monkey are
somewhat variable. Inspection of data for individual animals reveals
12
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changes in body weight of as great as 10% on successive 4-week weighings;
these changes may be either loss or gain. Analysis of variance showed
significant (p < 0.05) differences among the seven groups at all time
intervals except the fourth (Weeks 13 through 16); but corresponding
differences among the six HTO groups were apparent only during the first
time interval (Weeks 1 through 4). Further, no dose-response relation-
ship was apparent between body weight and HTO level among the six HTO
groups. Thus, although the control group weighed more than the groups
receiving HTO after the first 8 weeks, this relationship probably was
not associated with the HTO, unless some phenomenon independent of the
concentration of HTO occurred. For example, one possibility is that HTO
administered within the specified range of concentration has a taste that
is less acceptable than that of ordinary tap water. However, three other
factors distinguished the control group from the six HTO groups: (1) the
control pens were on the end of the two rows of seven pens in which
inseminated animals were housed; (2) HTO-receiving animals drank fluid
stored in polyethylene containers whereas the controls drank directly
from the tap-water pipes; (3) although the drinking valves were identical
for all seven groups, the valves supplying water for the controls were
about 25 cm above the pen floor, whereas those for the HTO groups were
about 100 cm above the floor. The decision not to use the polyethelene
tanks for controls was based on physical arrangements: tanks on the
outside of the end pens would have interfered markedly with access to
the corridors through which personnel and equipment moved. The height
difference was thought to be unimportant, since no decrements in the
agility or activity of the monkeys associated with HTO were observed.
Nonetheless, exploring the palatability of HTO as compared with that of
ordinary tap water would be of interest in a future study on this
species.
The body weight data for full-term pregnant females (Table 5) were
subjected to analysis of variance, and no differences (p < 0.05) among
the means for the seven groups were apparent. For evaluation of the
effects of full-term pregnancy on body weight, the individual data were
adjusted so that birth was allotted to Week 22, and the pooled (across
13
-------
dose groups) data were compared with the pooled postinsemination data
for the animals that did not deliver or abort; Figure 1 shows these
results. Because only a few full-term animals were weighed during the
first 3 weeks of pregnancy, the apparently lower mean weights during
that period for full-term mothers may not be significant. Clearly, from
about the sixth week, the mean body weight for full-term animals exceeded
that for the remaining animals, reaching values about 16% higher from
Week 15 until birth. Because of variability, however, increase in body
weight does not appear to be a useful criterion of pregnancy, since by
the time that significant weight excess would be reliably apparent
(Week 15), palpation and/or visible abdominal enlargement already would
have established pregnancy.
Births
Table 6 shows the time distribution of the 46 full-term progeny.
Although comparable numbers of inseminated females were in each of the
dose groups, the number of full-term progeny varied markedly, with no
consistent dose dependency discernible. The sex distribution among all
46 progeny was not remarkable; but all 4 of the Group 4 progeny were
male, and all 8 of the Group 7 progeny were female. Median and mean
times from insemination to birth were consistent among all groups. Thus,
these data indicate no clear effect of HTO administration on apparent
gestation period, sex distribution among progeny, or proportion of
inseminated animals delivering full-term progeny.
. Precise data for estimating the gestation period in the Bolivian
squirrel monkey are not available in the literature. In the present study,
however, the individual breeding histories are known, and they provide
a means for evaluating the range of possible gestation periods for each
of the animals. They also provide evidence for evaluating the accuracy
of the insemination criteria used and some information about breeding
behavior. Table 7 shows these data. Single entries in the first column
*
Completeness of hair covering and easy eyelid retraction were the
criteria establishing full-term status.
14
-------
900
800
I-
HI
Q
O
m
LU
5
700 -
600
I I I I I I I
NO DELIVERY OR FRANK ABORTION
y+2 S.E
i» MEAN
-L-2 S.E.
8 10 12
WEEKS POST - CONCEPTION
adjusted
FIGURE 1 COMPARISON OF POOLED BODY WEIGHT DATA FOR FULL-TERM PREGNANT FEMALES
WITH THOSE FOR THE REMAINING INSEMINATED ANIMALS
-------
identify animals that were bred continuously for the specified number of
days before insemination. Composite numbers indicate the number of days
for each segment of intermittent breeding, with the numbers in parenthe-
ses indicating the number of days between breeding periods. Thus, each
of the individual days postinsemination has a possible range equal to
the total number of days over which breeding could have occurred.
The best estimate of the minimum observed gestation period is
141 days because it is based on a viable full-term delivery for a female
bred for only 1 day. The last two animals in Table 7, although having
postinsemination times of 136 and 87 days, had possible maximum gestation
periods of 151 and 157 days. The data indicate that, at least in these
two cases, an earlier insemination apparently was not detected, but they
also indicate that either the observed insemination was in error or that
insemination occurred in already pregnant females. The latter situation
has been observed by Clewe,s who reported that copulation may occur as
late as the fourth month of pregnancy. All the remaining 44 animals had
observed gestational periods of 141 to 159 days, with a mean of 152.5
days and a standard deviation of 3.9 days. Gestation periods for the
Colombian squirrel monkey have been reported to range from 168 to
172 days.6 Our data indicate that the best estimate of the gestation
period for the Bolivian squirrel monkey is 141 to 158 days. The criteria
of insemination used in the present study apparently were valid, because
with the two exceptions notedall but one (159 days) of the observed
gestation periods were within the range of 141 to 158 days for the
animals bred for 1 day.
Concentrations of HTO in Body Water
Beginning with the week ending 6/4/76, all animals receiving HTO
in drinking water were placed in metabolism cages for 1 day each week
for collection of urine specimens for determining the body water content
of HTO and evaluating pregnancy. Animals were placed in the cages between
2 and 3 p.m. and were removed between 10 and 11 a.m. the following morning.
Two groups were placed in the metabolism cages each day Monday through
16
-------
Wednesday, and the seventh group (controls) was placed in the metabolism
cages on Thursday. The cages were washed, dried, and prepared for the
next groups of animals during the interval between 11 a.m. and 2 p.m.
While in the metabolism cages, the animals received HTO ad libitum from
the same water stock provided for them in their home pens. Initially,
food pellets were supplied; but, after the first few collections, we
found that the urine samples were contaminated extensively with food
debris, so food was withheld during the period in the metabolism cage.
Beginning with the week of 8/13/76, the schedule was changed to
a 4-week cycle for placing animals in metabolism cages, with two groups
being put in the cages each week for 3 weeks, and the seventh group (the
controls) being put in them during the fourth week. No other aspects
of the procedure were changed.
Urine excreted by the monkeys was collected in stainless-steel trays,
from which it drained into wide-mouth glass jars. After the monkeys were
removed in the morning, aliquots of the urine were decanted from the
jars into capped plastic tubes for assay of HTO content, and a second
aliquot was decanted for pregnancy assay. The results of the radio-
activity assay were tabulated by animal identification number and week of
collection. Usable urine specimens could not be obtained from all animals
at each collection time. In some cases, the volume of urine was so small
that it dried or was adsorbed on debris in the collecting tray; in a few
cases, the animal appeared not to have urinated during the confinement
in the metabolism cage. Because of the large number of animals being
handled, and the policy of keeping handling of potentially pregnant
animals to a minimum, the lack of urine specimens in a given week from
some of the animals was accepted as unavoidable, and no attempt was made
to collect additional specimens.
Table 8 summarizes results of the urine assays for HTO in animals
receiving HTO in the drinking water. The data are mean concentrations of
HTO in all animals of each group, listed by week of study. HTO concentra-
tions were lower during the first 6 weeks of the study, before the final
adjustment of HTO concentration in the drinking water. During the period
17
-------
after the final adjustment, the overall concentration of HTO in the urine
of each group was of the order of 80 to 90% of that originally intended
by the experimental protocol. However, the mean concentrations of HTO in
urine varied greatly from one week to the next in all groups, and the
standard deviations of concentration within each group for each week were
high, ranging up to as much as 40% of the mean.
Table 9 shows the data for assays of urine from the control animals
(Group 1). These animals were not given HTO but were supplied with
drinking water from drinking valves attached to the building water supply.
They were housed in the same building as the experimental animals.
Beginning with the fourth week of the study, HTO was detected in the
urine of these animals, and from 7/16 through 11/26 the overall concen-
tration of HTO in the urine was of the order of 9 to 10% of that in the
urine of Group 2 animals. The only possible source of HTO for these
animals was the HTO in the ambient air in the animal compound; this HTO
presumably came from expiration and from evaporation of water and urine
from the other pens in the compound.
Tables 10 through 16 summarize the individual measurements of HTO
concentration in urine for all the animals that delivered full-term
infants in Groups 1 through 7, respectively. Blanks in the tables indi-
cate weeks during which samples of urine were not obtained from the
individual animals.
We examined the data on concentration of HTO in urine to determine
whether pregnancy had any effect on the HTO concentration in body water.
The method used was to compute an overall mean and standard deviation for
all animals that delivered infants (including not-full-term infants) and
for all other animals in each experimental group except Group 1. Table 17
summarizes the results of the analysis. In five of the six groups, the
HTO concentration was slightly higher in pregnant than in nonpregnant
monkeys, ranging from 3 to 15%. The overall difference between pregnant
and nonpregnant monkeys was 4.8%, and the results suggest the influence
of pregnancy on the water balance but are of doubtful significance.
A more refined analysis of the data, focusing on particular stages of
pregnancy, might show stronger and more significant differences.
18
-------
The concentrations of HTO in urine were generally proportional to
those in the drinking water, but. the variability was very high and the
concentrations were inconsistent from week to week. Figure 2 illustrates
the weekly variation in urine-to-drinking water ratios for Groups 2 and 7.
The concentration ratios ranged from 0.46 to 0.68 and from 0.34 to 0.72
for Groups 7 and 2, respectively, and no systematic correlation could be
established between the two groups. Standard deviations (shown for
Group 7 only) were high. The results indicate that procedural physical
or physiological factors other than concentration of HTO in drinking water
strongly influenced the concentration of HTO in urine and body water.
The overall mean ratio of urine HTO concentration to that in drinking
water was determined after completion of the experiment by computing the
linear regression of the mean HTO concentrations in urine for all the
groups at all weeks (Table 8) on the corresponding HTO concentrations in
drinking water (Table 2). The completed regression analysis gave the
equation:
Urinary HTO = (0.010 ± 0.034) + (0.539 ± 0.018) (HTO in drinking water)
The regression line extrapolated to zero at zero drinking water and
showed that the overall average urinary-to-drinking water ratio for HTO
was 0.539. This value is considerably less than that expected on the
basis of studies with rats.7
Examination of the standard deviations of HTO concentration in various
groups indicated that they tended to be proportional to the means of the
groups. On the basis of this observation, the overall standard deviation
for concentration of HTO in urine was estimated by computing the linear
regression of the standard deviation on the mean for all the groups at
all weeks of study. The resulting line extrapolated to zero standard
deviation at zero mean HTO concentration and showed that the average
standard deviation for concentration of HTO in urine was 25.2% of the
mean. By using a similar procedure for successive duplicate samples of
drinking water, we found that the standard deviation for the concentra-
tion of HTO in drinking water was 7.3% of the mean. The standard
19
-------
Ni
O
7/2
7/30
8/27 9/24 10/22
WEEK OF URINE COLLECTION
11/19
12/17
1/14
FIGURE 2 RELATIONSHIP BETWEEN URINE AND DRINKING WATER CONCENTRATIONS OF HTO IN ADULT
FEMALE SQUIRREL MONKEYS OF GROUPS 2 AND 7
-------
deviation for HTO concentration in drinking water represents the vari-
ability sources included in preparing and mixing the drinking water and
determining its HTO content. The standard deviation for HTO concentra-
tion in urine represents all sources of variability. By subtracting
the square of drinking water standard deviation from the square of urine
standard deviation and by taking the square root of the result, we can
estimate the standard deviation for the urinary concentration of HTO,
exclusive of that contributed by the variability of the drinking water.
The value of this reduced standard deviation was 24.1% of the mean and
represented the variability of urinary concentration of HTO, as measured
in this study, resulting from various causes including variations in
animal individuality, feeding and drinking behavior, physiological state,
current food and water consumption, and metabolic rate.
Assay of Radioactivity in Tissues
Samples of selected organs taken from the neonate monkeys at necropsy
were kept frozen in plastic vials. Before homogenization, these samples
were weighed on an analytical balance. The tissues were homogenized in
an all-glass homogenizer with at least three volumes (w/v) of distilled
water. The final volume of each homogenate was recorded so that the volume
could be related to the initial wet weight of the tissue. Quadruplicate
aliquots (0.1 ml) of each homogenate were pipetted into polyethylene vials.
The homogenate was decolorized by the addition of 0.1 ml of 30% hydrogen
peroxide. After the samples had stood at ambient temperature for 15 to
20 minutes, 10 ml of scintillator fluid was added to each of two samples
for immediate radioactivity assay. The other two samples were placed in
an oven (temperature, 45 to 60° C) for overnight evaporation of moisture.
The residue was treated with 1.0 ml of ethanol and dried again in the
oven. The radioactivity in the dried samples was counted after addition
of scintillator fluid, in the same manner as described above.
*
Duall, Kontes Glass.
21
-------
As shown in Tables 18 and 19, respectively, the counting data were
expressed as microcuries per gram of wet tissue before and after removal
of the moisture. The difference between these two sets of data is
attributable to tritiated water in the tissue, and the radioactivity
content in tissues after dehydration is taken to represent tritium fixed
in the tissue by incorporation into tissue protein, carbohydrate, fat,
or nucleic acid.
The dependency of organ tissue radioactivity (wet) on the HTO concen-
tration in the drinking water, apparent from Tables 18 and 19, was
subjected to linear regression analysis. We found a significant (p < 0.05)
dependence of tissue radioactivity on HTO concentration and that the
relationship fitted (p < 0.05) the linear regression model for each of the
11 organs and/or tissues. The calculated slopes and Y-intercept (esti-
mated radioactivity at zero drinking water HTO concentration) are summar-
ized in Table 20. The 95% confidence limits for the estimated zero
concentration of HTO in the drinking water all include zero radioactivity.
This indicates a linear relationship with no threshold; that is, no HTO
concentration is apparent in drinking water where no radioactivity is
discernible in the tissues.
Gross and Microscopic Evaluations of Progeny
Gross and histopathologic evaluations of the neonate squirrel monkeys
were conducted according to the protocol specified in the contract with
three exceptions: (1) omission of most of the SMAC-20 blood chemistries
because of our inability to obtain adequate volumes of blood, (2) approxi-
mately one-half of the whole fixed brains were sent to EPA for future
special detailed neuropathologic evaluation, and (3) parts of some brains
and" all ovaries and eyes were sent to the Lawrence Livermore Laboratory
for future special evaluation. The necropsy procedure is outlined in
Schedule 1. Forty-one full-term viable progeny, five full-term stillborn
progeny, and four aborted fetuses were necropsied and examined. Formalin-
fixed sections were processed to H&E slides, with the exceptions noted
above.
22
-------
Schedule 1
SQUIRREL MONKEY NECROPSY PROCEDURE
Prenecropsy Sequence
Clinical, behavior, activity
Bleed from heart:
1 ml, CBC, Vacutainer
1 ml, Rad Count, Vacutainer
1-2 ml+ of Serum SMAC 20 (glass vial)
External examination for defects
Measurements
Weights, body and organ
Necropsy Sequence
Brain*
Pituitary
Eyes
Nasal cavity
Salivary gland and node
Larynx, trachea, esophagus
Skin and breast
Sternum
Thymus
Heart*
Lung
Diaphragm
Spleen*
Kidney*
Adrenal*
Ovaries/testes
Uterus/accessory gland
Urocyst (dilate)
Liver*
Stomach
Intestines
Mesenteric node
Sciatic nerve and muscle
Femur, crush for marrow
touch preparation
1/2 Brain
1/2 Liver
Radiation Count, Vial Number
2
Muscle
1/2 Heart
Skin
1/2 Placenta
1 Kidney
1 Adrenal
1/2 Stomach
< 1/2 Spleen
1" Duodenum
Weigh these organs.
23
-------
In five cases, gross lesions were observed at necropsy. One full-
term viable infant had hemorrhage in the brain in the meninges, focally
congested lungs, and an umbilical prolapse. Cerebral hemorrhage was
noted in one full-term stillborn infant, and hemoperitoneum was observed
in another. One aborted fetus had lacerations and contusions of the
right foot and left occiput, and another had a contused frontal area on
the skull. Most likely, these observations are related to postnatal
trauma, particularly in the case of the latter four.
Of the six cases of frank abortion, congestion, extramedullary
hematopoiesis, and lung atelectasis were the only microscopic observations.
Table 21 lists the locations of the lesions observed in the 46 full-term
viable and stillborn animals. Congestion was the diagnosis in over two-
thirds of the lesions, and about one-fourth of the lesions observed were
in the lung. No relationship between frequency of occurrence of lesions
and HTO dose was apparent, with one exception. Unlike the rest of the
neonate tissues examined histologically (including testes), the ovaries
of the female neonates were clearly affected by HTO. Illustrative slides
from animals of Groups 1, 5, and 7 are shown in Figures 3, 4, and 5,
respectively. In the control animal (Figure 3), many large oocytes are
visible, each surrounded by inconspicuous follicular cells, the latter
encompassed by scant connective tissue. External to the capsule is
a single row of cuboidal cells comprising the germinal epithelium. In
the neonates from the HTO groups, particularly at the highest dose level
(Figure 5), markedly fewer oocytes and many more follicular cells are
apparent.
Even on this subjective basis, the squirrel monkey oocyte population
appears to be remarkably sensitive to HTO exposure during gestation on
the basis of the body water concentration of HTO. For the mouse, exposure
to about 3.0 yCi/ml (body water) from conception through 14 days of age
results in an oocyte survival of about 34%. In the present study, animals
of Group 7 had a mean body water HTO level of about 3 yCi/ml, and the
oocyte survival in that group clearly was much less than 30%. The
apparent dose-response relationship observed in this study is being
24
-------
25x
FIGURE 3
'v 200x
LOW- AND HIGH-MAGNIFICATION PHOTOMICRO-
GRAPHS OF A GROUP 1 NEONATE OVARY
1!
-------
25x
200x
FIGURE 4 LOW- AND HIGH-MAGNIFICATION PHOTOMICRO-
GRAPHS OF A GROUP 5 NEONATE OVARY
26
-------
25x
200x
FIGURE 5 LOW- AND HIGH-MAGNIFICATION PHOTOMICRO-
GRAPHS OF A GROUP 7 NEONATE OVARY
2
-------
quantified by Dr. Dobson, who is evaluating the neonate ovaries from this
project using a quantitative procedure.8'9 When these data become avail-
able, the relative sensitivity of the squirrel monkey as compared with
other species can be assessed more meaningfully. Also, additional quanti-
tative evaluations of the retinas from all of the neonates and of the
brains from some will be attempted under similar collaborative
arrangements.
Finally, several adult female squirrel monkeys (from Groups 1 and
4 through 7) have been unilaterally ovariectomized, and the ovaries have
been fixed in a more appropriate fixative (Bouin's) for future quantitative
evaluation. This effort is being undertaken to provide information on
the ovaries of the HTO-exposed adults comparable to that for the neonates,
because studies in rodents have indicated that the oocyte count can be
reduced even with postnatal irradiation.8
Table 22 summarizes selected body dimensions, body weight, and the
weights of selected organs by dose group. No relationship between dose
of HTO and any of the 13 other parameters is apparent. However, a differ-
ence in dimensions and weight between male and female full-term progeny
was apparent. The same data tabulated according to sex are shown in
Tables 23 and 24. Analysis of variance indicated a significant (p < 0.025)
difference between the body weights of female progeny and those of male
progeny. Although not evaluated statistically, the values for each of
the other 12 parameters were greater for males than for females, consistent
with the body weight relationship.
Hematology and Serology
Inspection of the individual hematologic data revealed no apparent
differences between dose groups, and the data were pooled for statistical
description shown in Table 25. Values for RBC, Hgb, Hct, MCV, MCH, and
MCHC in the present study are in extremely close agreement with those
recently reported for neonate squirrel monkeys of Colombian origin.10
The latter report gives appreciably higher mean values than those shown
in Table 25 for WBC (11.07 vs 6.75) and for the percentage of neutrophils
28
-------
(77 vs 66.8), but the variability for these parameters appears relatively
large for both sets of data. The present data confirm the findings of
Ausman et al.,10 for the neonate squirrel monkey:11"13 elevated Hgb, Hct,
and RBC count, and macrocytes (MCV).
In only five cases was sufficient blood available for serology.
Table 26 shows these data. No conclusions about dose response are
possible. Compared with adult squirrel monkeys,10-12 neonates had some-
what higher values for calcium and cholesterol and markedly lower
globulin and carbon dioxide values. Corresponding adult values for
creatinine, triglycerides, and iron are not available, and apparent
differences in the alkaline phosphatase and SGPT methods prevent compari-
son for those parameters. Present values for protein appear comparable
to the mean neonate value of 5.46 g% reported by Ausman et al.,10 but the
present values for albumin appear somewhat higher than the mean of
3.08 g% they reported.
29
-------
Table 1.
SCHEDULED HTO VALUES
Group
Number
1
2
3
4
5
6
7
Desired
Body HTO
(uCi/ml)
0
0.05
0.2
0.5
1.0
2.0
4.0
Initial
Injection
(mCi)
0
0.023
0.091
0.23
0.46
0.91
1.82
Drinking Water HTO
(yd/ml)
Initial
0
0.060
0.240
0.600
1.200
2.410
4.820
Final
0
0.072
0.290
0.720
1.450
2.900
5.800
30
-------
Table 2. MEAN ASSAYED HTO DRINKING WATER ACTIVITY
(Microcuries per Milliliter)
N
Mean
S.D.
N
Mean
S.D.
N
Mean
S.D.
Date
6/7/76
6/11
6/18
6/25
7/2
7/9
7/16
7/23
7/30
8/6
8/13
8/20
8/27
9/3
9/10
9/17
9/24
10/1
10/8
10/15
10/22
10/29
11/5
11/12
11/19
11/26
12/3
12/10
12/17
12/24
12/31
1/7/77
1/14
2
0.048
0.045
0.047
0.040
4
0.045
0.004,
0.064
1
0.064
_. _
0.071
0.081
0.077
0.071
__
0.094
0.065
0.081
0.056
0.097
0.094
0.078
0.077
0.070
0.074
0.068
0.077
0.082
0.072
0.076
0.068
0.075
0.072
22
0.076
0.010
3
0.186
0.196
0.181
0.149
4
0.178
0.020
0.247
1
0.247
0.321
0.357
0.320
0.316
__
0.299
0.258
0.303
0.211
0.306
0.300
0.299
0.288
0.286
0.265
0.275
0.262
0.266
0.280
0.262
0.283
0.264
0.279
0.263
23
0.285
0.030
Group
4
0.483
0.472
0.469
0.370
4
0.448
0.053
0.598
1
0.598
m.M
0.637
0.667
0.704
0.838
__
0.682
0.716
0.714
0.572
0.732
0.700
0.768
0.740
0.714
0.660
0.740
0.820
0.719
0.761
0.712
0.767
0.722
0.766
22
0.720
0.058
5
0.938
0.938
0.915
0.824
4
0.904
0.054
1.189
1
1.189
__
1.463
1.378
__
1.462
1.282
1.397
1.190
1.386
1.417
1.414
1.380
1.427
1.324
1.386
1.346
1.440
1.517
1.413
1.494
1.467
1.539
20
1.406
0.081
6
1.856
1.932
1.847
1.637
4
1.818
0.127
2.347
2.333
2
2.340
0.010
__
2.874
2.894
2.873
3.431
__
3.016
2.723
2.906
2.584
2.896
2.816
2.822
2.766
2.952
2.600
2.838
2.876
3.004
3.130
2.935
3.131
3.006
3.171
22
2.920
0.189
7
4.013
3.607
3.581
3.225
4
3.606
0.322
4.763
4.545
2
4.654
0.154
5.742
5.714
5.787
5.749
__
5.930
5.431
5.849
5.760
5.723
5.730
5.974
5.690
5.862
5.663
6.332
5.738
5.881
6.210
5.940
6.312
5.960
6.130
22
5.868
0.220
31
-------
Table 3. PREGNANCY EVALUATION DATA FOR INSEMINATED FEMALE SQUIRREL MONKEYS
RECEIVING HTO IN DRINKING WATER
u>
to
Group
1
2
3
4
5
6
7
Total
No.
Insemi-
nated
43
38
38
41
38
39
41
277
No.
Evalu-
ated
37
35
33
36
36
36
33
246
No.
Positive
15
18
12
19
20
22
15
121
No. of
Full-Term
Deliveries
6
7
3
4
7
9
7
43*
No. of
Frank
Abortions
1
0
3
1
3
0
0
8*
No. of
Resorp-
tions
8
11
6
14
10
13
8
70
Percent
Resorp-
tion
53
61
50
74
50
59
53
58
Percent
Pregnant
40
51
36
53
56
61
46
49
R
Three full-term deliveries and two frank abortions occurred in animals not evaluated
for pregnancy.
-------
Table 4. BODY WEIGHTS OF INSEMINATED FEMALE SQUIRREL MONKEYS
NOT PROVIDING FULL-TERM DELIVERIES
OR EVIDENCE OF FRANK ABORTION
(Grams)
Weeks Post Insemination
Group Item
1 N
Mean
S.D.
2 N
Mean
S.D.
3 N
Mean
S.D.
4 N
Mean
S.D.
5 N
Mean
S.D.
6 N
Mean
S.D.
7 N
Mean
S.D.
Ito 4
13
635
26
6
657
60
7
703
55
10
659
41
12
699
60
13
635
32
13
672
59
5 to 8
49
722
66
44
695
48
46
715
56
58
704
55
48
718
63
54
702
41
58
692
43
9 to 12
45
754
73
39
711
55
40
723
59
36
708
51
31
730
63
34
725
49
33
708
55
13 to 16
31
757
86
28
720
57
30
735
73
31
733
59
22
747
72
26
743
63
28
722
47
17 to 20
37
771
78
29
728
61
29
729
66
29
735
64
28
736
64
26
739
67
27
713
46
21 to 24
33
787
72
24
724
60
24
764
82
23
752
77
21
741
75
25
750
60
27
723
54
33
-------
Table 5. BODY WEIGHTS OF FEMALE SQUIRREL MONKEYS
DELIVERING FULL-TERM PROGENY
(Grams)
Group Item
1 N
Mean
S.D.
2 N
Mean
S.D.
3 N
Mean
S.D.
4 N
Mean
S.D.
5 N
Mean
S.D.
6 N
Mean
S.D.
7 N
Mean
S.D.
Ito4
2
704
11
2
583
45
_
M ,
_
___
1
622
5 to 8
7
754
66
12
709
39
9
718
39
6
716
30
11
754
72
17
736
62
14
759
51
Weeks of
9 to 12
8
787
79
9
735
40
3
727
64
6
737
38
10
765
65
11
773
59
12
792
56
Pregnancy
13 to 16
6
879
102
8
789
60
4
787
85
4
798
91
7
848
83
10
838
84
8
853
76
17 to 20
6
914
125'
8
857
64
4
822
86
4
844
107
7
893
89
9
903
80
8
919
80
21 to 22
2
867
2
3
862
32
2
868
68
2
789
44
4
896
90
5
916
103
1
879
34
-------
Table 6. TIME DISTRIBUTION OF FULL-TERM BIRTHS
M = Male; F = Female
Group
Days Post- v
insemination ~ ,
87 F*
136 F
141
144
145 Mf
148
149 M
151
152 F MF M
153 F F
154 M FfF MM
155 M M
156 M F Ff MM
157
158
159
Total 6844
Median 152.5 154.0 154.0 155.6
Mean 149.3 153.1 154.2 154.8
ATI
AJ-.L
567
1*
1
M 1
FfM 2
M 2
F 1
1
FF MMF F 6
F 5
M FFFF 7
MM 7
2
F 6
F 1
FF 2
M 1
7 9 8 46
151.0 151.0 153.0 153.0
151.4 150.4 153.4 152.1
Excluded from mean.
t
Stillborn.
35
-------
Table 7. ESTIMATES OF GESTATION PERIOD FOR THE SQUIRREL MONKEY
Days of No. of Individual Days Post insemination to Birth
Breeding Animals
1
2
3
4
5
6
7
8
9
10
11
13
14
-15
26
2(1)1
2(13)1(4)1
2(6)7
37(32)1
7
5
1
2
4
4
3
6
3
2
1
1
1
1
1
1
1
1
1
141 151 152 153 153 154 158
151 153 153 154 154
151
152 158
152 152 155 156
149 151 151 153
153 154 156
144 144 153 154 154 155
154 156 157
145 152
156
159
148
145
156
156
151
136
87
36
-------
Table 8. MEAN HTO CONCENTRATIONS IN URINE OF INSEMINATED ADULT FEMALE SQUIRREL MONKEYS
(Microcuries per Milliliter)
N «= Number of Samples
Wc-<-k
T'.nd i ng
h/4/76
h/l 1
h/18
6/25
7/2
7/9
7/16
7/23
7/30
H/6
8/13
H/20
H/27
9/10
9/17
9/24
10/8
1 ()/ 1 r>
10/22
H/r>
11/12
11/19
12/3
1 'il 1(1
12/17
12/31
1/7/77
1/14
N
2
7
18
13
24
30
28
27
24
30
22
27
18
25
22
2
Group
Mean'
0.032
0.015
0.025
0.026
0.024
0.032
0.047
0.050
0.049
0.054
0.042
0.045
0.063
0.030
0.035
0.027
2
S.D.
0.002
0.006
0.006
0.006
0.005
0.011
0.015
0.007
0.009
0.013
0.005
0.009
O.OL1
0.010
0.006
0.005
N
2
9
17
11
26
29
23
22
29
28
19
26
27
15
20
3
Group
Mean
0.138
0.093
0.099
0.093
0.112
0.121
0.151
O.J93
0.199
0.181
0.161
0.165
0.164
0. 127
0. 130
0.064
3
S.D.
0.018
0.033
0.024
0.011
0.034
0.048
0.045
0.038
0.041
0.044
0.037
0.038
0.039
0.022
0.023
0.056
N
3
7
14
19
21
27
*
23
27
28
32
32
34
24
22
6
Group
Mean
0.316
0.185
0.252
0.227
0.290
0.298
0.357
0.382
0.444
0.293
0.361
0.335
0.384
0.330
0.288
4
S.D.
0.046
0.054
0.037
0.100
0.042
0.068
0.086
0.075
0.080
0.134
0.103
0.091
0.156
0.067
0.082
N
4
7
14
14
27
26
*
25
27
34
35
33
31
19
17
7
Group
Mean
0.695
0.565
0.556
0.523
0.512
0.520
0.897
0.850
0.833
0.690
0.631
0.801
0.471t
0.667
0.722
5
S.D.
0.122
0.197
0.059
0.081
0.130
0.112
0.186
0.165
0.131
0.276
0.179
0.193
0.143
0.211
0.113
N
3
7
15
13
25
28
31
32
27
20
31
34
33
28
23
8
Group
Mean
1.175
1.268
1.128
0.971
1.132
1.175
1.534
1.832
1.832
1.954
1.884
1.326
1.446
1.315
1.485
1.346
6
S.D.
0.202
0.303
0.234
0.208
0.329
0.432
0.340
0.277
0.419
0.291
0.486
0.431
0.289
0.299
0.385
0.419
N
2
5
16
16
22
28
20
25
22
18
33
33
32
34
20
7
Group
Mean
2.641
2.344
2.427
2.032
2.427
2.290
3.267
3.124
2.987
2.945
3.318
2.707
2.785
2.877
3.486
3.232
7
S.D.
0.086
0.234
0.182
0.386
0.557
0.845 '
1.095
0.810
0.835
0.634
0.819
0.726
0.725
0.602
0.900
0.863
Samp I ir-; lost by .ice I dent.
Puss i l> U- c out ami nation wi l h ra in water.
-------
Table 9. CONCENTRATIONS OF HTO
IN URINE OF CONTROL ANIMALS
NOT RECEIVING HTO IN DRINKING WATER
(Microcuries per Milliliter)
Week
Ending
6/4/76
6/11
6/18
6/25
7/2
7/9
7/16
7/23/76
7/30/76
8/6
9/3
10/1
10/29
11/26
12/24
1/12/77
No. of
Animals
Tested
3
8
14
19
26
29
25
29
28
34
23
31
14
23
5
5
Mean
0
0
0
0.00116
0.00019
0.00052
0.00108
0.00483
0.00764
0.00324
0.00461
0.00416
0.00464
0.00335
0.00180
0.00140
S.D.
0.00112
0.00049
0.00078
0.00064
0.00275
0.00296
0.00148
0.00244
0.00267
0.00139
0.00147
0.00045
0.00089
38
-------
Table 10. URINARY CONCENTRATION OF HTO
IN GROUP 1 ADULT SQUIRREL MONKEYS DELIVERING FULL-TERM PROGENY
(Microcuries per Milliliter)
Date
(1976)
6/4
6/18
6/25
7/2
7/9
7/16
7/23
7/30
8/6
9/3
10/1
10/29
11/26
Days
*
Mean
S.D.
542
< 0.001
< 0.001
0.001
0.001
0.003
0.008
0.003
0.002
0.002
0.003
136
0.0024
0.0005
546
0.001
< 0.001
0.001
< 0.001
0.001
< 0.001
0.003
0.005
0.002
0.002
0.006
145
0.0020
0.0004
Animal
641
< 0.001
0.001
< 0.001
0.001
< 0.001
0.003
0.009
0.001
0.003
0.002
0.002
156
0.0026
0.0006
Number
800 813
< 0.001
< 0.001
0.002
0.006
0.010
0.006 0.005
0.008
0.005 0.006
0.007 0.005
0.004
153 152
0.0044 0.0056
0.0010 0.0012
852
0.001
0.013
0.004
0.004
0.008
0.006
0.005
0.002
154
0.0050
0.0011
Weighted according to the number of days of pregnancy during the
periods 5/29 - 6/25, 6/26 - 7/9, and 7/10 - delivery. Dash indicates
no satisfactory sample was obtained; blank indicates animal was not
yet inseminated or was postpartum.
39
-------
Table 11. URINARY CONCENTRATION OF HTO
IN GROUP 2 ADULT SQUIRREL MONKEYS DELIVERING FULL-TERM PROGENY
(Microcuries per Milliliter)
Date
(1976)
6/4
6/11
6/18
6/25
7/2
7/9
7/16
7/23
7/30
8/6
8/27
9/24
10/22
1.1/19
Days
Mean
S.D.
Animal
503 545
0.
0.
0.
0.
0.
0.
0.
0.
0.049
0.045 0.
0.043 0.
0.
87
0.046 0.
0.010 0.
022
025
035
027
027
036
046
050
044
077
077
152
049
Oil
656
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
014
027
025
021
038
049
045
041
046
031
038
059
155
038
008
685
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
027
017
044
057
059
064
068
045
051
066
152
054
012
Number
692
0.033
0.029
0.027
0.049
0.055
0.060
0.040
0.066
0.043
0.053
0.056
149
0.050
0.011
0
0
0
0
0
0
0
0
0
0
0
717
.032
.040
.054
.063
.063
.067
.041
.064
.043
154
.053
.012
737
0.029
0.035
0.038
0.052
0.056
0.069
0.031
0.049
154
0.047
0.010
819
0.022
0.047
0.052
0.042
0.039
0.047
0.047
156
0.045
0.010
Weighted according to the number of days of pregnancy during the periods
5/29 - 6/25, 6/26 - 7/9, and 7/10 - delivery. Dash indicates no satis-
factory sample was obtained; blank indicates animal was not yet
inseminated or was postpartum.
40
-------
Table 12. URINARY CONCENTRATION OF HTO
IN GROUP 3 ADULT SQUIRREL MONKEYS
DELIVERING FULL-TERM PROGENY
(Microcuries per Milliliter)
Date
(1976)
6/11
6/18
6/25
7/2
7/9
7/16
7/23
7/30
8/6
8/27
9/24
10/22
11/19
Days
*
Mean
S.D.
719
0.085
0.087
0.103
0.098
0.298
0.169
0.111
0.135
0.123
0.197
ISA
0.157
0.035
Animal Number
747 764
0.118 0.098
0.123 0.038
0.124
0.163
0.222
0.102
0.187
0.167 0.191
0.184 0.174
0.083
154 156
0.145 0.171
0.032 0.038
805
0.097
0.146
0.052
0.169
0.181
0.187
0.179
0.207
0.164
153
0.156
0.034
Weighted according to the number of days of pregnancy during
the periods 5/29 - 6/25, 6/26 - 7/9, and 7/10 - delivery.
Dash indicates no satisfactory sample was obtained; blank
indicates animal was not yet inseminated or was postpartum.
41
-------
Table 13. URINARY CONCENTRATION OF HTO
IN GROUP 4 ADULT SQUIRREL MONKEYS
DELIVERING FULL-TERM PROGENY
(Microcuries per Milliliter)
Date
(1976)
6/4
6/11
6/18
6/25
7/2
7/9
7/23
7/30
8/6
8/20
9/17
10/15
11/12
Days
Mean
S.D.
640
0.280
0.274
0.306
0.281
0.386
0.149
0.415
0.522
156
0.341
0.075
Animal
686
0.366
0.158
0.243
0.314
0.273
0.324
0.279
0.332
0.403
0.415
0.411
156
0.338
0.074
Number
688
0.087
0.177
0.033
0.285
0.252
0.367
0.306
0.282
0.243
155
0.251
0.055
702
0.250
0.042
0.215
0.353
0.381
0.454
0.435
0.537
0.328
0.526
152
0.387
0.085
*
Weighted according to the number of days of pregnancy during
the periods 5/29 - 6/25, 6/26 - 7/9, and 7/10 - delivery.
Dash indicates no satisfactory sample was obtained; blank
indicates animal was not yet inseminated or was postpartum.
42
-------
Table 14. URINARY CONCENTRATION OF HTO
IN GROUP 5 ADULT SQUIRREL MONKEYS DELIVERING FULL-TERM PROGENY
(Microcuries per Milliliter)
Date
(1976)
6/4
6/11
6/18
6/25
7/2
7/9
7/23
7/30
8/6
8/20
9/17
10/15
11/12
Days
« *
Mean
S.D.
Animal Number
500
0.871
0.907
0.591
0.173
0.443
0.764
1.088
0.880
0.131
0.920
0.958
159
0.748
0.164
575
0.497
0.363
0.505
0.556
0.826
0.986
0.896
0.839
0.812
0.776
154
0.766
0.168
693
0.656
0.611
0.637
0.653
0.558
0.682
0.675
1.083
1.010
0.754
0.811
0.854
157
0.803
0.177
696
0.477
0.418
0.583
1.461
0.737
0.643
0.687
0.734
0.790
151
0.784
0.172
732
0.675
0.336
0.939
0.911
0.462
0.649
0.694
0.834
0.506
144
0.690
0.152
740
0.487
0.921
0.911
0.732
0.966
0.874
0.980
0.621
144
0.667
0.147
761
0.777
0.563
0.905
1.451
0.758
1.239
0.236
151
0.795
0.175
Weighted according to the number of days of pregnancy during the
periods 5/29 - 6/25, 6/26 - 7/9, and 7/10 - delivery. Dash indicates
no satisfactory sample was obtained; blank indicates animal was not
yet inseminated or was postpartum.
43
-------
Table 15. URINARY CONCENTRATION OF HTO
IN GROUP 6 ADULT SQUIRREL MONKEYS DELIVERING FULL-TERM PROGENY
(Microcuries per Milliliter)
Date
(1976) 515
6/4
6/11 1.165
6/18 0.978
6/25
7/2 0.976
7/9
7/16 1.444
7/23
7/30
8/6
8/13 1.587
9/10 1.697
10/8 1.077
11/5
Days 151
Animal Number
549
0.
0.
0.
0.
0.
0.
1.
1.
1.
1.
2.
1.
2.
942
833
965
968
985
954
357
761
767
844
101
811
007
151
556
0.961
0.910
1.300
1.943
1.763
2.273
2.164
2.204
2.200
1.641
1.530
1.831
152
561
1.
1.
0.
1.
1.
2.
2.
2.
1.
1.
1.
128
036
998
840
671
583
163
150
622
900
989
154
1
0
1
1
1
1
1
1
1
1
1
1
628
.063
.664
.333
.174
.540
.668
.795
.676
.388
.377
.611
.456
151
689
1.015
0.300
0.756
1.344
1.751
1.647
2.041
1.554
1.424
1.537
153
711
1.492
1.244
1.567
2.035
2.132
2.235
1.551
1.505
1.666
1.490
156
718
1.353
0.885
1.055
1.445
2.076
1.598
1.832
1.858
1.734
145
791
0.496
2.168
1.824
1.454
2.071
1.960
1.912
1.407
141
Mean 1.352 1.566 1.827 1.855 1.452 1.472 1.717 1.618 1.578
S.D. 0.297 0.345 0.402 0.408 0.319 0.324 0.378 0.356 .0.347
Weighted according to the number of days of pregnancy during the periods
5/29 - 6/25, 6/26 - 7/9, and 7/10 - delivery. Dash indicates no satisfactory
sample was obtained; blank indicates animal was not yet inseminated or was
postpartum.
44
-------
Table 16. URINARY CONCENTRATION OF HTO
IN GROUP 7 ADULT SQUIRREL MONKEYS DELIVERING FULL-TERM PROGENY
(Microcuries per Milllliter)
Date
(1976)
6/4
6/11
6/18
6/25
7/2
7/9
7/16
7/23
7/30
8/6
8/13
9/10
10/8
11/5
12/3
12/31
Days
Mean*
S.D.
508
2.269
2.728
2.561
2.323
2.383
3.810
3.025
2.789
2.332
3.236
3.965
148
3.004
0.661
585
2.500
2.380
2.883
2.582
3.440
3.823
4.961
3.322
2.699
3.581
3.976
3.860
153
3.428
0.754
606
2.607
1.481
2.102
2.368
2.593
3.213
2.331
2.898
3.586
3.518
3.690
153
2.925
0.644
Animal
661
2.580
1.975
2.313
1.302
2.323
3.499
3.129
2.904
2.859
4.044
2.826
3.417
3.444
153
2.971
0.654
Number
722
2.654
1.903
2.448
2.324
5.011
2.644
2.545
2.740
3.072
2.388
2.951
158
2.928
0.644
763 798
1.755
1.445 1.474
4.831
3.316
1.881 2.565
2.152
3.651 2.975
3.570 3.612
3.282 1.782
3.527 3.581
158 153
2.990 3.055
0.658 0.672
884
4.019
2.237
2.794
3.162
3.792
4.343
151
3.391
0.746
Weighted according to the number of days of pregnancy during the periods
5/29 - 6/25, 6/26 - 7/9, and 7/10 - delivery. Dash indicates no satis-
factory sample was obtained; blank indicates animal was not yet-
inseminated or was postpartum.
45
-------
Table 17. ANALYSIS OF MEAN* HTO CONCENTRATION IN URINE
OF PREGNANT AND NONPREGNANT FEMALE SQUIRREL MONKEYS
Group
2 34567
Nonpregnant
Mean HTO (uCi/ml) 0.0391 0.149 0.349 0.666 1.470 2.776
Number of samples 245 251 250 215 257 240
Pregnant
Mean HTO (yCi/ml) 0.0447 0.154 0.313 0.718 1.537 3.018
Number of samples 69 52 51 92 92 86
Difference
yCi/ml +0.0056 +0.005 -0.036 +0.052 +0.067 +0.242
Percent +14.6 +3.4 -10.3 +7.8 +4.6 +8.7
Pooled S.D. 0.0148 0.0496 0.106 0.219 0.447 0.785
t for difference 2.74 0.63 3.23 1.91 1.23 2.44
Unweighted, microcuries per milliliter.
46
-------
Table 18. TISSUE RADIOACTIVITY OF NEONATE SO.UIRSEL MONKEYS
(Microcurles per Gram of Wet Tissue)
N
Mean
S.D.
N
Mean
S.D.
N
Mean
S.D.
N
Mean
S.D.
N
Mean
S.D.
N
Mean
S.D.
N
Mean
S.D.
Animal
Number
1.542
1.546
1.641
1.800
1.813
1.852
2.503
2.544
2.656
2.685
2.692
2.717
2.737
2.819
3.719
3.747
3.764
3.805
4.640
4.686
4.688
4.702
5.500
5.575
5.693
5.696
5.732
5.740
5.761
6.515
6.439
6.556
6.561
6.628
6.689
6.711
6.718
6.791
7.508
7.585
7.606
7.661
7.722
7.763
7.798
7.884
Brain
0.007
0.011
0.001
3
0.006
0.005
0.054
0.047
0.063
0.041
4
0.051
0.009
0.142
0.170
2
0.156
0.020
0.507
0.624
0.497
3
0.542
0.070
0.765
0.954
1.096
3
0.938
0.166
1.647
1.903
1.692
1.747
1.356
1.439
6
1.675
0.244
3.718
3.713
2.870
2.511
4
3.202
0.609
Liver
0.006
0.010
0.003
0.002
0.003
0.001
6
0.004
0.003
0.051
0.044
0.049
0.042
0.045
0.041
0.038
0.036
8
0.043
0.005
0.133
0.157
0.144
0.127
4
0.140
0.013
0.537
0.469
0.404
0.369
4
0.445
0.074
0.771
0.746
0.875
0.983
0.603
0.701
0.634
7
0.759
0.133
1.273
1.412
1.584
1.710
0.973
1.377
1.482
1.406
1.531
9
1.416
0.209
3.310
3.379
1.331
2.487
2.584
3.373
3.051
3.339
8
2.856
0.712
Kid
Q.016
0.010
0.006
0.003
0.003
0.001
6
0.006
0.005
0.042
0.048
0.046
0.036
0.044
0.029
0.040
0.032
8
0.039
0.006
0.136
0.114
0.153
0.135
4
0.134
0.016
0.395
0.504
0.399
0.402
4
0.425
0.053
0.674
0.819
0.925
0.815
0.507
0.736
0.616
7
0.726
0.140
1.518
1.261
1.045
1.945
0.707
1.512
1.337
1.411
1.509
9
1.360
0.344
4.165
3.563
1.092
2.521
1.695
3.427
3.133
3.153
8
2.843
1.019
Adr
0.007
0.011
0.005
0.006
0.006
0.001
6
0.006
0.003
0.045
0.033
0.038
0.034
0.036
0.040
0.039
0.027
8
0.036
0.005
0.140
0.115
0.133
0.118
4
0.126
0.012
0.373
0.483
0.390
0.231
4
0.369
0.104
0.536
0.554
0.911
0.746
0.442
0.623
0.612
7
0.632
0.153
1.118
0.941
0.695
1.383
0.643
0.997
0.969
1.104
1.432
9
1.031
0.267
3.796
2.676
0.744
1.848
0.805
2.132
1.565
3.127
8
2.086
1.077
Spl
0.011
0.012
0.001
0.007
0.001
5
0.006
0.005
0.030
0.040
0.036
0.019
0.029
0.042
0.046
0.033
8
0.034
0.008
0.130
0.096
0.121
0.117
4
0.116
0.014
0.374
0.426
0.371
0.222
4
0.348
0.087-*
0.403
0.535
1.045
0.662
0.838
0.606
0.543
7
0.661
0.215
0.947
0.396
1.209
1.142
0.822
1.108
0.622
7
0.892
0.299
3.140
2.196
0.866
1.518
0.490
3.076
2.375
1.634
8
1.911
0.964
GIT
0.008
0.003
0.002
0.004
0.001
5
0.003
0.002
0.051
0.028
0.038
0.025
0.033
0.032
0.035
0.026
8
0.033
0.008
0.076
0.101
0.116
0.123
4
0.104
0.020
0.294
0.440
0.270
3
0.335
0.092
0.559
0.571
0.908
0.606
0.166
0.727
0.547
7
0.583
0.224
0.848
0.501
1.213
0.519
1.043
1.111
0.999
1.413
8
0.955
0.320
2.289
0.560
1.983
0.803
2.747
2.54:
2.097
7
1.860
0.847
Plac
0.012
0.004
2
0.008
0.005
0.029
0.044
0.028
0.042
4
0.035
0.008
0.106
0.143
2
0.124
0.026
0.344
1
0.344
0.960
0.811
0.710
0.627
0.507
5
0.722
0.172
1.478
0.632
2
1.055
0.598
1.413
1
1.-13
Skin
0.003
0.008
0.001
0.001
0.001
0.001
6
0.002
0.002
0.035
0.049
0.047
0.041
0.040
0.045
0.046
0.034
8
0.042
0.005
0.163
0.316
0.186
0.136
4
0.200
0.079
0.350
0.267
0.469
3
0.362
0.102
0.942
0.815
0.790
0.845
0.619
0.762
0.797
7
0.795
0.097
0.973
1.172
1.800
1.909
1.144
1.260
1.555
1.300
1.482
9
1.399
0.312
5.045
3.258
2.920
3.264
3.198
2.908
:.30S
7
3.212
O.S-9
Blood
0.002
0.001
0.001
0.001
4
0.001
0.000
0.044
0.041
0.046
0.043
0.024
0.021
6
0.036
0.011
0.099
0.078
0.125
3
0.100
0.023
0.397
0.265
0.390
3
0.350
0.074
0.696
0.588
0.788
0.603
0.410
0.645
6
0.621
0.126
1.202
1.245
1.347
0.986
1.324
1.195
1.450
1.267
8
1.252
0.136
3.581
3.467
2.945
2.832
2.598
2.094
3.362
7
2.982
0.532
V.us
0.009
0.006
0.003
0.005
0.001
3
0.004
0.003
0.046
0.040
0.044
0.036
0.036
0.039
0.027
7
0.038
0.006
0.140
0.124
0.143
0.105
4
0.128
0.017
0.414
0.422
0.363
0.455
4
0.414
0.038
0.819
O.S26
0.715
0.842
0.671
0.686
6
0.760
0.076
1.428
1.329
0.520
1.695
0.382
0.856
1.252
1.572
1.490
9
1.169
0.471
3.495
0.828
2.510
1.453
2.365
2.900
1.502
7
2.150
' 0.930
Heart
0.006
0.014
0.003
0.005
0.008
0.001
6
0.006
0.004
0.029
0.04i
0.038
0.032
0.039
0.040
0.038
0.039
8
0.037
0.004
0.124
0.086
0.121
0.123
4
0.114
0.018
0.353
0.429
0.405
0.292
4
0.370
0.061
0.524
0.721
0.751
0.788
0.353
0.620
0.628
7
0.626
0.150
1.257
1.254
0.180
1.427
0.499
1.137
1.076
1.366
1.483
9
1.075
0.443
i.041
2.456
1.133
2.262
0.583
2.876
2.513
2.890
8
2.3.4
1.074
In this computerized format (also used in subsequent tables), the integer is the group number and the remainder
Is the animal number.
47
-------
Tabl. 19.
TISSUE RADIOACTIVm OF NEONAIE SQUIRREL MONKEYS AFTER DEHYDRATION
(Mlcrocuries oer Cram of Wet Tissue)
N
Mean
N
Mean
S.D.
N
Mean
S.D.
X
Mean
S.D.
X
Mean
S.D.
X
Mean
S.D.
X
MiMn
S.D.
Animal
Number
1.542
1.546
1.641
1.800
1.813
1.852
2.503
2.545
2.656
2.685
2.692
2.717
2.737
2.819
3.719
3.747
3.764
3.805
4.640
4.686
4.688
4.702
5.500
i.575
5.693
5.696
5.732
5.710
5.761
A.M5
«i.5i9
A. 556
6.561
*>.«!l
6.h89
1.711
6.7IS
fc.791
7.i08
7.58>
:.wb
7.6M
7.722
7.76)
7.79H
7.384
Brain
0.001
0.001
0.001
3
0.001
0.001
0.003
0.004
0.001
4
0.002
0.001
0.003
0.006
2
0.004
0.002
0.009
0.011
0.011
3
0.010
0.001
0.026
0.030
0.030
3
0.029
n.oo:
1.039
0.031
0.070
0.05:
n.051
0.033
6
n.oi6
0.014
0.092
0.073
0.179
0.050
4
0.098
n.056
Liver
0.001
0.001
0.001
0.001
0.001
0.001
6
0.001
0.001
0.003
0.007
0.001
0.001
0.001
0.001
0.001
8
0.002
0.002
0.001
0.003
0.003
0.002
4
0.002
0.001
0.001
0.005
0.004
0.008
4
0.004
0.003
0.012
0.016
0.011
0.013
0.018
0.002
0.014
7
0.012
0.003
0.013
0.021
0.022
0.029
0.048
0.015
i).023
0.019
0.039
9
0.025
0.011
0.064
0.031
0.121
0.019
0.010
0.055
0.050
0.039
8
0.059
0.032
Kid
0.001
0.001
0.001
0.001
0.001
0.001
6
0.001
0.001
0.004
0.007
0.001
0.001
0.001
0.001
0.001
8
0.002
0.002
0.001
0.003
0.002
0.002
4
0.002
0.001
0.003
0.003
0.007
3
0.004
0.002
0.006
0.014
0.009
0.007
0.013
0.002
0.010
7
0.008
0.003
0.012
0.022
0.032
0.037
0.034
0.018
0.028
0.018
0.028
9
0.025
0.008
0.051
0.051
0.089
0.023
0.069
0.078
0.0i7
0.062
8
0.059
0.020
Adr
0.001
0.001
0.001
0.001
0.001
0.001
6
0.001
0.001
0.010
0.014
0.001
0.001
o.ooi
0.001
0.001
8
0.003
0.005
0.001
0.006
0.001
0.003
4
0.003
0.002
0.003
0.008
0.005
0.020
4
0.009
0.008
0.024
0.061
0.022
0.014
0.020
0.002
0.023
7
0.023
0.018
0.031
0.075
0.058
0.022
0.056
0.025
0.057
0.045
0.066
9
0.048
0.018
0.120
0.052
0.220
0.128
0.105
0.247
0.188
0.071
8
0,111
0.069
Spl
0.001
0.001
0.001
0.001
0.001
5
0.001
0.001
0.016
0.015
0.001
0.001
0.001
0.001
0.001
8
0.001
0.006
0.001
0.005
0.001
0.003
i
0.002
0.001
0.005
0.001
0.002
0.011
4
0.004
0.004
0.004
0.020
0.009
0.010
0.015
0.011
0.016
7
0.012
0.005
0.018
0.065
0.016
0.006
0.030
0.028
0.025
7
0.026
0.018
0.050
0.041
0.119
0.012
0.139
0.091
1.070
0.049
8
0.071
0.042
GIT
0.001
0.001
0.001
0.001
0.001
5
0.001
0.004
0.007
0.008
0.001
0.001
0.001
0.001
0.001
8
0.003
0.002
0.001
0.003
o.ooi
0.003
1
0.002
0.001
0.004
0.004
0.008
3
0.005
0.002
0.012
0.018
0.008
0.006
0.008
0.003
0.012
7
0.009
0.004
0.031
0.029
0.010
0.021
0.020
0.023
0.016
0.029
8
0.022
0.007
0.040
0.100
0.022
0.048
0.078
0.053
0.027
7
0.052
0.027
Plac
0.001
0.001
2
0.001
0.001
0.003
0.001
0.001
4
0.001
0.001
0.001
0.002
2
0.001
0.000
0.001
1
0.001
0.049
0.005
0.002
0.005
0.023
5
0.016
0.019
0.020
0.014
2
0.017
0.004
0.031
1
0.031
Skin
0.001
0.001
0.001
0.001
0.001
0.001
6
0.001
0.002
0.002
0.005
0.001
0.001
0.001
0.001
0.001
8
0.001
0.001
0.006
0.010
0.007
0.004
4
0.006
0.002
0.016
0.004
0.019
3
0.013
0.008
0.023
0.02!
0.012
0.032
0.013
0.042
0.036
7
0.026
0.011
0.021
0.012
0.036
0.069
0.031
0.060
0.034
0.034
0.036
9
0.011
0.011
0.294
0.063
0.120
0.089
C.066
0.132
0.128
7
0.127
0.078
Blood
0.001
0.001
0.001
0.001
4
0.001
0.001
o.ooi
0.001
0.001
0.001
0.001
6
0.001
0.001
0.001
0.001
3
0.001
0.001
o.ooi
0.001
3
0.001
0.002
0.002
0.002
0.002
0.001
0.001
6
0.001
o.ooo
0.002
0.004
0.004
0.006
0.001
0.008
0.004
0.005
0.003
9
0.004
0.001
0.008
0.007
0.007
0.006
0.013
0.009
O.Olh
7
0.009
0.003
Mus
0.001
0.001
0.001
0.001
0.001
5
0.001
0.007
0.005
0.001
0.001
0.001
0.001
0.001
7
0.002
0.002
0.003
0.003
0.002
0.002
4
0.002
0.001
0.004
0.003
0.003
0.011
u
0.005
0.004
0.016
P. 016
0.013
0.010
0.014
0.012
6
0.012
0.002
0.011
0.029
0.031
0.034
0.031
0.006
0.016
0.021
0.027
9
0.023
0.010
0.026
0.061
0.022
0.060
0.070
0.052
0.022
7
0.044
0.020
Heart
0.001
0.001
0.001
0.001
0.001
0.001
6
0.001
0.001
0.005
0.002
0.001
0.001
0.001
0.001
0.001
8
0.001
0.001
o.ooi
0.004
0.001
0.002
1
0.002
0.001
0.003
0.003
0.001
0.007
4
0.004
0.002
0.007
o.oi:
0.008
0.005
0.016
0.006
o.on
7
0.009
O.OOi
0.009
0.015
0.0:5
0.030
0.023
0.007
0.0:2
O.C11
0.021
9
0.018
0.008
0.068
0.025
O.J172
0.024
o.n63
O.f'56
T.053
0.116
8
0.059
0.029
48
-------
Table 20. LINEAR REGRESSION ANALYSIS OF TISSUE RADIOACTIVITY
IN THE NEONATE SQUIRREL MONKEY
Brain
Liver
Kidney
Adrenal
Spleen
GIT
Placenta
Skin
Blood
Muscle
Heart
Slope
0.531
0.487
0.489
0.373
0.322
0.318
0.386
0.538
0.513
0.383
0.405
*
LCL
0.486
0.454
0.441
0.321
0.270
0.271
0.305
0.492
0.479
0.332
0.352
UCLf
0.575
0.521
0.538
0.425
0.375
0.364
0.466
0.584
0.546
0.434
0.458
Y- intercept
0.066
0.020
0.002
0.022
0.045
0.046
0.043
- 0.023
- 0.067
0.059
0.000
*
LCL
- 0.023
- 0.049
- 0.097
- 0.084
- 0.063
- 0.046
- 0.094
- 0.115
- 0.137
- 0.043
- 0.108
UCL1"
0.157
0.090
0.101
0.130
0.153
0.139
0.181
0.069
0.002
0.163
0.108
LCL = Lower 95% confidence limit.
UCL = Upper 95% confidence limit.
49
-------
Table 21. DISTRIBUTION OF HISTOLOGIC LESIONS
AMONG FULL-TERM NEONATE SQUIRREL MONKEYS
Viable
(N=41)
Still
(N=5)
All
Group
1
2
3
4
5
6
7
Sum
1
2
3
4
5
6
7
Sum
1
2
3
4
5
6
7
Adr
1
0
1
1
0
0
0
3
0
2
-
1
0
-
-
3
1
2
1
2
0
0
0
Kid
0
0
1
0
0
0
0
1
0
1
-
1
0
-
-
2
0
1
1
1
0
0
0
Liv
0
0
1
0
1
0
0
2
1
-
1
0
-
-
2
0
1
1
1
1
0
0
Lung
2
1
3
0
1
1
2
10
1
0
-
0
1
-
-
2
3
1
3
0
2
1
2
Brain
0
0
0
0
0
0
0
0
1
-
0
0
-
-
1
0
1
0
0
0
0
0
Pane
0
0
1
0
0
0
0
1
-
-
-
0
-
-
0
0
0
1
0
0
0
0
Jej
0
0
0
0
0
0
0
0
0
0
-
1
0
-
-
1
0
0
0
1
0
6
0
Pit
0
0
0
0
1
0
1
2
0
-
1
-
-
-
1
0
0
0
1
- 1
0
1
Spl
0
0
1
0
0
0
0
1
0
0
-
1
-
-
-
1
0
0
1
1
0
0
0
Tes
0
0
0
0
0
0
-
0
-
-
1
-
-
-
1
0
0
0
1
0
0
-
Thym
0
0
0
0
0
6
0
0
0
1
-
0
0
-
-
1
0
1
0
0
0
0
0
Skin
0 .
0
0
0
1
0
0
1
0
0
-
0
0
-
-
0
0
0
0
0
1
0
0
All
3
1
8
1
4
1
3
21
1
6
-
7
1
-
-
15
4
7
8
7
5
I
3
Sum 6 3 4 12 1 1 1 3 2 1 1 1 36
50
-------
Table 22. BODY DIMENSIONS AND ORGAN WEIGHTS IN NEONATE SQUIRREL MONKEYS BY HTO CONCENTRATION
N
Mean
S.D.
N
Mean
S.D.
N
Mean
S.D.
N
Mean
S.D.
N
Mean
S.D.
N
Mean
S.D.
N
Mean
S.D.
Animal
Number
1.542
1.546
1.641
1.800
1.813
1.852
2.503
2.545
2.656
2.685
2.692
2.717
2.737
2.819
3.719
3.747
3.764
3.805
4.640
4.686
4.688
4.702
5.500
5.575
5.693
5.696
5.732
5.740
5.761
6.515
6.549
6.556
6.561
6.628
6.689
6.711
6.718
6.791
7.508
7.585
7.606
7.661
7.722
7.763
7.798
7.884
8
Head* (cm)
P-P
2.50
2.90
3.60
3.40
3.80
4.30
6
3.41
0.64
2.90
3.90
3.60
3.60
3.80
3.30
3.40
3.50
8
3.50
0.31
3.60
3.10
3.80
3.30
4
3.45
0.31
3.50
3.60
3.00
3.60
4
3.42
0.28
3.80
3.60
3.50
3.60
3.60
2.90
3.90
7
3.55
0.32
2.60
3.30
3.60
3.50
3.10
3.70
2.90
3.90
3.00
9
3.28
0.42
3.20
3.40
3.80
3.60
3.50
3.60
3.20
3.30
8
3.45
0.21
N-0
4.70
4.60
5.40
5.40
5.30
5.20
6
5.10
0.35
4.80
5.40
4.40
5.10
5.30
4.90
4.90
5.50
8
5.03
0.36
5.80
5.10
5.20
4.40
4
5.12
0.57
5.20
5.30
5.20
4.90
4
5.15
0.17
5.00
5.40
5.00
5.20
5.20
5.00
5.10
7
5.12
0.14
4.60
5.40
5.20
5.40
4.70
5.20
4.00
5.70
4.80
9
5.00
0.52
5.20
5.10
5.30
5.10
4.90
5.10
5.10
5.00
8
5.04
0.13
C-C
3.20
3.20
3.30
3.30
4.00
3.10
6
3.35
0.32
3.40
3.00
3.40
3.80
3.70
3.40
3.30
3.70
8
3.46
0.26
3.90
4.40
3.10
3.80
4
3.80
0.53
4.00
3.20
3.40
3.60
4
3.55
0.34
3.50
3.10
3.60
3.70
3.70
2.80
3.20
7
3.37
0.34
3.00
3.40
3.90
3.20
3.40
3.40
3.40
3.60
2.90
9
3.35
0.30
3.40
3.20
3.20
3.60
3.00
3.10
3.40
3.00
8
3.23
0.20
Body (cm)
Thor
2.40
2.70
2.90
2.80
2.80
2.60
6
2.70
0.17
2.40
2.80
2.80
2.60
2.90
2.40
2.60
2.70
8
2.65
0.18
2.70
2.40
2.80
2.50
4
2.60
0.18
2.90
3.00
2.80
2.70
4
2.85
0.13
2.80
3.10
2.40
2.70
2.60
1.80
2.80
7
2.60
0.41
2.50
2.80
2.50
3.10
2.50
. 2.90
2.70
2.80
1.50
9
2.58
0.45
2.60
2.90
2.90
3.10
2.40
3.00
2.70
1.60
8
2.65
0.48
Abdo
2.20
2.20
2.40
2.40
2.00
2.60
6
2.30
0.20
2.00
2.30'
2.70
1.90
2.30
1.80
2.10
2.20
8
2.16
0.28
2.10
1.90
2.50
2.00
4
2.12
0.26
2.20
2.90
2.40
2.10
4
2.40
0.35
2.20
2.50
2.10
2.20
3.20
1.60
2.30
7
2.30
0.48
2.30
2.60
2.00
2.70
2.10
2.20
2.10
2.20
1.90
9
2.23
0.26
2.00
2.50
2.50
2.40
2.10
2.40
2.20
2.10
8
2.27
0.19
Lngth
12.10
13.30
14.40
13.70
13.20
15.00
6
13.61
1.01
11.90
14.20
14.00
12.60
13.60
12.80
13.20
13.40
8
13.21
0.76
13.60
12.40
12.90
13.80
4
13.17
0.64
14.00
13.70
13.30
13.30
4
13.57
0.34
13.30
13.70
' 12.40
14.00
13.10
12.70
13.40
7
13.22
0.55
11.80
13.50
13.90
13.60
12.80
12.80
12.60
13.20
12.20
9
12.93
0.68
13.40
14.00
13.50
13.40
13.20
12.60
13.20
12.70
8
13.25
0.46
Body
82.00
91.20
127.00
110.00
94.00
129.00
6
105.53
19.61
77.00
126.00
121.00
95.00
108.00
80.00
102.00
101.00
8
101.25
17.43
127.00
84.00
99.00
103.00
4
103.25
17.82
116.00
119.00
106.40
96.00
f 4
109.35
10.39
105.00
114.00
97.00
112.00
98.00
98.00
117.00
7
105.85
8.47
94.00
111.90
107.00
125.00
89.00
110.00
88.00
126.00
80.00
9
103.43
16.53
95.70
114.00
103.00
124.00
103.00
113.00
105.00
88.20
8
105.74
11.20
Brain
11.00
8.70
14.42
15.06
14.34
16.01
6
13.25
2.80
13.30
16.20
15.70
14.52
15.69
14.16
14.78
14.64
8
14.87
0.94
15.00
13.75
14.68
14.44
4
14.46
0.53
16.25
16.10
15.20
14.51
4
15.51
0.81
15.40
16.50
14.85
13.85
15.00
14.10
15.84
7
15.07
0.93
14.40
13.60
15.24
15.74
14.70
14.54
13.93
16.67
13.68
9
14.72
1.01
14.80
13.50
15.24
15.20
13.05
14.00
14.74
12.40
8
14.11
1.04
Weight (g)
Hrt
0.45
0.90
0.66
0.56
0.56
0.98
6
0.68
0.20
0.42
0.76
0.70
0.56
0.64
0.42
0.67
0.55
8
0.59
0.13
0.80
0.47
0.57
0.58
4
0.60
0.13
0.88
0.91
0.70
0.58
4
0.77
0.16
0.65
0.75
0.65
0.69
0.70
0.75
0.74
7
0.70
0.04
0.60
0.80
0.74
0.76
0.60
0.62
0.52
0.68
0.60
9
0.65
0.09
0.50
0.95
0.59
0.66
0.68
0.67
0.47
0.50
8
0.63
0.16
Adr
0.12
0.13
0.19
0.18
0.12
0.15
6
0.15
0.03
0.14
0.20
0.20
0.12
0.18
0.17
0.22
0.10
8
0.17
0.04
0.25
0.22
0.14
0.34
4
0.23
0.08
0.26
0.40
0.20
0.17
4
0.25
0.10
0.11
0.20
0.25
0.20
0.40
0.50
0.35
7
0.28
0.13
0.20
0.25
0.16
0.20
0.30
0.13
0.14
0.11
0.30
9
0.19
0.07
0.15
0.17
0.13
0.10
0.12
0.10
0.10
0.10
8
0.11
0.02
Kid
0.65
0.75
0.90
0.76
0.53
0.89
6
0.74
0.14
0.36
0.94
0.90
0.55
0.65
0.42
0.61
0.70
8
0.64
0.21
0.97
0.44
0.57
0.68
4
0.66
0.22
0.74
1.00
0.90
0.62
4
0.81
0.16
0.67
0.85
0.60
0.81
0.70
0.85
0.71
7
0.74
0.09
0.65
0.70
0.61
0.70
0.70
0.65
0.47
0.74
0.60
9
0.64
0.08
0.70
0.66
0.48
0.86
0.55
0.55
0.70
0.50
8
0,63
0.12
Llv
2.10
2.25
4.36
3.64
3.01
3.17
6
3.08
0.85
4.30
3.59
4.70
2.72
3.62
2.48
2.65
3.22
8
3.41
0.80
3.46
2.84
3.57
4.91
4
3.69
0.87
6.05
4.10
3.30
2.86
4
4.07
1.41
3.10
3.35
3.60
3.73
2.70
4.20
3.34
7
3.43
0.47
3.20
2.70
3.06
4.88
2.50
3.73
2.68
3.56
2.30
9
3.17
0.79
2.65
3.76
3.76
3.66
2.82
2.25
3.28
2.50
8
3.08
0.60
Spl
0.10
0.11
0.13
0.13
0.15
0.13
6
0.13
0.03
0.11
0.16
0.10
0.10
0.17
0.12
0.13
0.18
8
0.13
0.03
0.15
0.15
0.13
0.16
4
0.14
0.01
0.18
0.30
0.20
0.15
4
0.20
0.06
0.13
0.25
0.20
0.14
0.25
0.30
0.11
7
0.19
0.07
0.20
0.04
0.11
0.12
0.20
0.14
0.15
0.10
0.20
9
0.14
0.05
0.15
0.13
0.09
0.17
0.09
0.10
0.14
0.10
8
0.12
0.03
P-P - parietal width; N-0 = nose to occiput; C-C = chin to crown.
51
-------
Table 23. BODY DIMENSIONS AND ORGAN WEIGHTS OF MALE NEONATE SQUIRREL MONKEYS
Ui
to
Animal
Number
1.546
1.641
1.852
2.545
2.656
2.692
3.719
3.747
4.640
4.686
4.688
4.702
5.500
5.575
5.732
6.549
6.561
6.628
6.689
6.718
6.791
N
Mean
S.D.
Head (cm)
P-P
2.90
3.60
4.30
3.90
3.60
3.80
3.60
3.10
3.50
3.60
3.00
3.60
3.80
3.60
3.60
3.30
3.50
3.10
3.70
3.90
3.00
21
3.52
0.35
N-0
4.60
5.40
5.20
5.40
4.40
5.30
5.80
5.10
5.20
5.30
5.20
4.90
5.00
5.40
5.20
5.40
5.40
4.70
5.20
5.70
4.80
21
5.17
0.34
C-C
3.20
3.30
3.10
3.00
3.40
3.70
3.90
4.40
4.00
3.20
3.40
3.60
3.50
3.10
3.70
3.40
3.20
3.40
3.40
3.60
2.90
21
3.44
0.35
Thor
2.70
2.90
2.60
2.80
2.80
2.90
2.70
2.40
2.90
3.00
2.80
2.70
2.80
3.10
2.60
2.80
3.10
2.50
2.90
2.80
1.50
21
2.63
0.53
Body (cm)
Abdo
2.20
2.40
2.60
2.30
2.70
2.30
2.10
1.90
2.20
2.90
2.40
2.10
2.20
2.50
3.20
2.60
2.70
2.10
2.20
2.20
1.90
21
2.73
0.33
Lngth
13.30
14.40
15.00
14.20
14.00
13.60
13.60
12.40
14.00
13.70
13.30
13.30
13.30
13.70
13.10
13.50
13.60
12.80
12.80
13.20
12.20
21
13.47
0.64
Body
91.20
127.00
129.00
126.00
121.00
108.00
127.00
84.00
116.00
119.00
106.40
96.00
105.00
114.00
98.00
111.90
125.00
89.00
110.00
126.00
80.00
21
109.97
15.30
Brain
8.70f
14.42
16.01
16.20
15-. 70
15.69
15.00
13.75
16.25
16.10
15.20
14.51
15.40
16.50
15.00
13.60
15.74
14.70
14.54
16.67
13.68
21
14.92
1.69
Weight (g)
Hrt
0.90
0.66
0.98
0.76
0.70
0.64
0.80
0.47
0.88
0.91
0.70
0.58
0.65
0.75
0.70
0.80
0.76
0.60
0.62
0.68
0.60
21
0.72
0.12
Adr
0.13
0.19
0.15
0.20
0.20
0.18
0.25
0.22
0.26
0.40
0.20
0.17
0.11
0.20
0.40
0,25
0.20
0.30
0.13
0.11
0.30
21
0.21
0.08
Kid
0.75
0.90
0.89
0.94
0.90
0.65
0.97
0.44
0.74
1.00
0.90
0.62
0.67
0.85
0.70
0.70
0.70
0.70
0.65
0.74
0.60
21
0.76
0.14
Liv
2.25
4.36
3.17
3.59
4.70
3.62
3.46
2.84
6.05
4.10
3.30
2.86
3.10
3.35
2.70
2.70
4.88
2.50
3.73
3.56
2.30
21
3.48
0.93
Spl
0.11
0.13
0.18
0.16
0.10
0.17
0.15
0.15
0.18
0.30
0.20
0.15
0.13
0.25
0.25
0.04
0.12
0.20
0.14
0.10
0.20
21
0.16
0.05
P-P = parietal width; N-0 = nose to occiput; C-C = chin to crown.
This animal was found dead on pen floor.
fracture were observed.
At.necropsy, contusions and herniation of the brain through a skull
-------
Table 24. BODY DIMENSIONS AND ORGAN WEIGHTS OF FEMALE NEONATE SQUIRREL MONKEYS
Animal
1.542
1.800
1.8]3
2.503
2.685
2.717
2.737
2.819
3.764
3.805
5.693
5.696
5.740
5.761
6.515
6.556
6.711
7 . 508
7.585
7.606
7.661
7.722
7.763
7.798
7.884
N
Mc-an
S.D.
Head (cm)
P-P
2.50
3.40
3.80
2.90
3.60
3.30
3.40
3.50
3.80
3.30
3.50
3.60
2.90
3.90
2.60
3.60
2.90
3.20
3.40
3.80
3.60
3.50
3 . 60
3.30
3.30
25
3.36
0.36
N-0
4.70
5.40
5.30
4.80
5.10
4.90
4.90
5.50
5.20
4.40
5.00
5.20
5.00
5.10
4.60
5.20
4.00
5.20
5.10
5.30
5.10
4.90
5.10
5.10
5.00
25
5.00
0.32
C-C
3.20
3.30
4.00
3.40
3.80
3.40
3.30
3.70
3.10
3.80
3.60
3.70
2.80
3.20
3.00
3.90
3.40
3.40
3.20
3.20
3.60
3.00
3.10
3.40
3.00
25
3.38
0.31
Body (cm)
Thor
2.40
2.80
2.80
2.40
2.60
2.40
2.60
2.70
2.80
2.50
2.40
2.70
1.80
2.80
2.50
2.50
2.70
2.60
2.90
2.90
3.10
2.40
3.00
2.70
1.60
25
2.58
0.33
Abdo
2.20
2.40
2.00
2.00
1.90
1.80
2.10
2.20
2.50
2.00
2.10
2.20
1.60
2.30
2.30
2.00
2.10
2.00
2.50
2.50
2.40
2.10
2.40
2.20
2.10
25
2.15
0.22
Lngth
12.10
13.70
13.20
11.90
12.60
12.80
13.20
13.40
12.90
13.80
12.40
14.00
12.70
13.40
11.80
13.90
12.60
13.40
14.00
13.50
13.40
13.20
12.60
13.20
12.70
25
13.05
0.62
Body
82.00
110.00
94.00
77.00
95.00
80.00
102.00
101.00
99.00
103.00
97.00
112.00
98.00
117.00
94.00
107.00
88.00
95.70
114.00
103.00
124.00
103.00
113.00
105.00
88.20
25
100.07
11.62
Brain
11.00
15.06
14.34
13.30
14.52
14.16
14.78
14.64
14.68
14.44
14.85
13.85
14.10
15.84
14.40
15.24
13.93
14.80
13.50
15.24
15.20
13.05
14.00
14.74
L2.40
25
14.24
1.01
Weight (g)
Hrt
0.45
0.56
0.56
0.42
0.56
0.42
0.67
0.55
0.57
0.58
0.65
0.69
0.75
0.74
0.60
0.74
0.52
0.50
0.95
0.59
0.66
0.68
0.67
0.47
0.50
25
0.60
0. 12
Adr
0.12
0.18
0.12
0.14
0.12
0.17
0.22
0.10
0.14
0.34
0.25
0.20
0.50
0.35
0.20
0.16
0.14
0.15
0.17
0.13
0.10
0.12
0.10
0.10
0.10
25
0.17
0.09
Kid
0.65
0.76
0.53
0.36
0.55
0.42
0.61
0.70
0.57
0.68
0.60
0.81
0.85
0.71
0.65
0.61
0.47
0.70
0.66
0.48
0.86
0.55
0.55
0.70
0.50
25
0.62
0.12
Liv
2.10
3.64
3.01
4.30
2.72
2.48
2.65
3.22
3.57
4.91
3.60
3.73
4.20
3.34
3.20
3.06
2.68
2.65
3.76
3.76
3.66
2.82
2.25
3.28
2.50
25
3.24
0.68
Spl
0.10
0.13
0.15
0.11
0.10
0.12
0.13
0.18
0.13
0.16
0.20
0.14
0.30
0.11
0.20
0.11
0.15
0.15
0.13
0.09
0.17
0.09
0.10
0.14
0.10
25
0 . 1 3
0 . 04
?-'' = parietal width; N-0 = nose to occiput; C-C = chin to crown.
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Table 25. HEMATOLOGIC PARAMETERS OF NEONATE SQUIRREL MONKEYS
Ui
Animal
Number
1.542
1.800
1.813
1.852
2.545
2.692
2.717
2.737
2.819
3.719
3.747
4.686
4.688
5.575
5.693
5.696
5.732
5.761
6.515
6.549
6.556
6.561
6.689
6.711
6.718
6.791
7.585
7.606
7.661
7.763
7.798
7.884
N
Mean
S.D.
RBC
(xlO~6)
4.70
6.45
5.88
5.61
5.54
5.40
6.19
6.41
3.76
6.74
5.88
7.28
6.06
5.55
5.93
6.21
6.11
6.53.
6.41
5.94
4.39
5.97
6.72
4.03
5.32
4.65
6.57
5.88
5.87
5.70
6.43
5.25
32.
5.79
0.80
Hgb
(g Z)
16.30
19.40
18.30
18.80
17.70
17.70
19.20
19.90
10.80
20.50
17.70
22.20
19.30
16.40
18.50
17.90
18.10
19.00
18.30
19.00
13.60
17.30
20.80
12.30
16.10
15.20
19.20
18.50
17.30
17.30
18.30
17.30
32
17.75
2.31
Hct
(%)
46.30
55.50
52.20
53.30
50.60
50.90
54.60
57.70
31.80
57.50
52.30
63.40
56.10
48.60
53.00
52.10
51.00
55.70
52.70
53.40
39.50
50.80
59.50
34.50
46.70
43.40
55.80
51.80
51.00
49.30
54.40
49.80
32
51.10
6.56
MCV
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Table 26. SEROLOGIC PARAMETERS OF NEONATE SQUIRREL MONKEYS
Animal Number
Glucose (mg%)
BUN (mg%)
Great inine (mg%)
Uric acid (mg%)
Na+ (meq/1)
K+ (meq/1)
C02 (meq/1)
Cl~ (meq/1)
Calcium (mg%)
Phosphorus (mg%)
Balance (5-7-8) (meq/1)
Cholesterol (mg%)
Triglycerides (mg%)
Bilirubin (Tot) (mg%)
SCOT (mU/ml)
SGPT (mU/ml)
LDM (mU/ml)
Alk. phos. (mU/ml)
Iron (Tot) (mcg%)
Protein (Tot) (g%)
Albumin (g%)
Globulin (g%)
A/G (g%)
Bilirubin (Dir) (mg%)
5.575
140
24
0.8
0
141
3.5
16
101
11.3
3.2
24
334
43
8.4
143
33
263
1410
69
4.9
4.4
0.5
8.80 -
0.2
6.515
302
26
1.1
0.7
146
5.6
17
96
11.0
4.6
33
313
32
13.6
152
3
342
1810
41
5.4
5.1
0.3
17.00
6.549
87
27
1.2
0.8
150
3.4
22
100
10.2
5.0
28
225
40
9.7
238
24
371
1870
54
4.6
4.3
0.3
14.33
7.585
216
35
1.2
1.7
148
5.6
15
107
11.8
5.0
26
269
246
9.4
372
46
1060
2490
65
5.4
4.9
0.5
9.80
7.661
138
18
0.9
0
146
4.1
14
109
10.2
4.8
23
331
13
8.7
96
14
415
1845
149
5.0
4.5
0.5
9.00
55
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REFERENCES
1. D. J. Kimeldorf and E. L. Hunt. Ionizing radiation and neural
development (Ch. 3). In Ionizing Radiation: Neural Function and
Behavior. Academic Press, New York, 1976.
2. U.S. Nuclear Regulatory Commission, Title 10, Code of Federal
Registry, Part 20, Appendix B, 30 May 75.
3. W. W. Tullner and R. Hertz. Chorionic gonadotrophin levels in the
Rhesus monkey during early pregnancy. Endocrinology 78, 204-207
(1966).
4. J. G. Wilson and R. Fradkin. Teratogeny in nonhuman primates, with
notes on breeding procedures in Macacca mulatta. Ann. N.Y. Acad.
Sci. 162, 267-277 (1969).
5. T. H. Clewe. Observations on reproduction of squirrel monkeys in
captivity. J. Reprod. Fertil. (Suppl.) £, 151-156 (1969).
6. C. M. Goss, L. T. Popejoy, J. L. Fusiler, and T. M. Smith.
Observations on the relationship between embryological development,
time of conception, and gestation (Ch. 6). In The Squirrel Monkey.
L. A. Rosenblum and R. W. Cooper (eds.), Academic Press, New York,
1968.
7. D. F. Cahill and C. L. Yuile. Tritium: Some effects of continuous
exposure in utero on mammalian development. Radiat. Res. 44, 727-
737 (1970).
8. R. L. Dobson and C. T. Kwan. The RBE of tritium radiation measured
in mouse oocytes: Increase at low exposure levels. Radiat. Res.
66, 615-625 (1976).
9. R. L. Dobson and M. F. Cooper. Tritium toxicity: Effects of low-
level 3HOH exposure on developing female germ cells in the mouse.
Radiat. Res. _5J3, 91-100 (1974).
10. L. M. Ausman, D. L. Gallina, K. C. Hayes, and D. M. Hegsted.
Hematological development of the infant squirrel monkey (_S_aimiri
sciureus). Folia Primatol. ^6, 292-300 (1976).
11. A. E. New. App. Base-line blood determinations of the squirrel
monkey (Saimirix sciureus). Appendix to The Squirrel Monkey.
L. A. Rosenblum and R. W. Cooper (eds.), Academic Press, New York,
1968.
56
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12. P. J. Manning, N.D.M. Lehner, M. A. Feldner, and B. C. Bullock.
Selected hematologic, serum chemical, and arterial blood gas
characteristics of squirrel monkeys (Saimiri sciureus). Lab.
Animal Care 3.9, 831-837 (1969).
13. C. M. Lang. The laboratory care and clinical management of
Saimiri (squirrel monkeys) (Ch. 14). In The Squirrel Monkey.
L. A. Rosenblum and R. W. Cooper (eds.), Academic Press, New York,
1968.
14. C. C. Middleton and J. Rosal. Weights and measurements of normal
squirrel monkeys. Lab. Animal Sci. 22, 583-585 (1972).
15. K. Capel-Edwards and D. E. Hall. Haemotological observations on
the squirrel monkey. Folia Primatol. 12, 142-160 (1970).
57
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TECHNICAL REPORT DATA
'Please read lnantcrions on the reverse before completing)
1. REPORT NO
EPA-600/1-78-004
2.
3. RECIPIENT'S ACCESSION NO.
4. TITLE AND SUBTITLE
EVALUATION OF NEONATE SQUIRREL MONKEYS RECEIVING
TRITIATED WATER THROUGHOUT GESTATION
5. REPORT DATE
January 1978
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
David C.L. Jones
8. PERFORMING ORGANIZATION REPORT NO.
9. PERFORMING ORGANIZATION NAME AND ADDRESS
Stanford Research Institute
Menlo Park, California 9.4025
10. PROGRAM ELEMENT NO.
1HA630
11. CONTRACT/GRANT NO.
68-02-2280
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
RTP.NC
13. TYPE OF REPORT AND PERIOD COVERED
14. SPONSORING AGENCY CODE
EPA 600/11
15. SUPPLEMENTARY NOTES
16. ABSTRACT
The effect of receiving tritiated water (HTO) throughout gestation on the
developing primate was assessed by administering HTO to adult female squirrel
monkeys (Saimiri sciureus) as the only source of drinking water beginning with the
day of insemination and continuing throughout pregnancy. For the control (tap
water), and six experimental groups, the mean urinary tritium concentrations in
females delivering full-term progeny were <0.004, 0.05, 0.16, 0.33, 0.75, 1.61,
and 3.09 yCi/ml. Positive bioassays for pregnancy were observed in about half of
277 inseminated females. Among pregnant females, the full-term delivery rate was
36%, the abortion rate was 7%, and the resorption rate was 58% with no discernible
effect of HTO administration on any of these parameters. The 46 full-term progeny
were evaluated within 2 days of birth. No effects of HTO administration were
observed in terms of gestation period (median 153 days, range 141-158 days), sex
distribution, body weight, body dimensions, selected organ weights, histology
(except gonads), or hematologic pattern. The number of primary oocytes in female
progeny decreased markedly within increasing concentrations of tritium. Specific
quantification of this effect and evaluation of the neonate testes is in progress.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.lDENTIFIERS/OPEN ENDED TERMS C. COSATI Field/Group
water-
tritium
health
monkeys
tritiated water
06 R, T
13. DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
19. SECURITY CLASS (ThisReport)
UNCLASSIFIED
21. NO. OF PAGES
65
20. SECURITY CLASS (Thispage)
UNCLASSIFIED
22. PRICE
EPA Form 2220-1 (9-73)
58
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