SWRHL-54r
ANIMAL INVESTIGATION PROGRAM BASELINE STUDIES:
SUPPLEMENTAL TEST SITE "C"
by
Bruce P. Hull, Jr., D. V. M. , and Stanley L. Cohen, D. V. M.
Animal Investigation Program
Bioenvironmental Research
Southwestern Radiological Health Laboratory
Department of Health, Education, and Welfare
Public Health Service
National Center for Radiological Health
September 13, 1968
This surveillance performed under a Memorandum of
Understanding (No. SF 54 373)
for the
U. S. ATOMIC ENERGY COMMISSION
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LEGAL NOTICE
This report -was prepared as an account of Government sponsored
•work. Neither the United States, nor the Atomic Energy Commission,
nor any person acting on behalf of the Commission:
A. makes any warranty or representation, expressed or implied,
with respect to the accuracy, completeness, or usefulness of the in-
formation contained in this report, or that the use of any information,
apparatus, method, or process disclosed in this report may not in-
fringe privately owned rights; or
B. assumes any liabilities with respect to the use of, or for damages
resulting from the use of any information, apparatus, method, or pro-
cess disclosed in this report.
As used in the above, "person acting on behalf of the Commission"
includes any employee or contractor of the Commission, or employee
of such contractor, to the extent that such employee or contractor of
the Commission, or employee of such contractor prepares, dissemin-
ates, or provides access to, any information pursuant to his employ-
ment or contract with the Commission, or his employment with such
contractor.
077
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SWRHL-54r
ANIMAL INVESTIGATION PROGRAM BASELINE STUDIES:
SUPPLEMENTAL TEST SITE "C"
by
Bruce P. Hull, Jr., D.V.M., and Stanley L. Cohen, D.V.M.
Animal Investigation Program
Bioenvironmental Research
Southwestern Radiological Health Laboratory
Department of Health, Education, and Welfare
Public Health Service
National Center for Radiological Health
September 13, 1968
This surveillance performed under a Memorandum of
Understanding (No. SF 54 373)
for the
U. S. ATOMIC ENERGY COMMISSION
-------
ABSTRACT
The Animal Investigation Progam (AIP) collected tissue samples from
two Mule deer and three beef cattle for radioanalysis and histo-
pathologic studies prior to the Faultless event. Pathologic lesions
present and detectable body burdens of selected radioisotopes were
comparable to those found in routine sacrifices.
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TABLE OF CONTENTS
ABSTRACT i
TABLE OF CONTENTS ii
LIST OF TABLES AND FIGURE iii
INTRODUCTION 1
PROCEDURES 2
RESULTS AND DISCUSSION--RADIOANALYSIS 4
RESULTS AND DISCUSSION—PATHOLOGY 5
SUMMARY 6
APPENDIX
DISTRIBUTION
11
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LIST OF TABLES AND FIGURE
Table A-l.
Table A-2.
Table A-3.
Table A-4.
Table A-5.
Table A-6.
Table A-7.
Table A-8.
Thyroid
Muscle
Rumen Contents
Abomasum Contents
Abomasum Tissue
Liver
Lung
Bone
7
7
'8
8
8
9
9
9
Figure 1.
Proposed Supplement Test Site and Morey Bench
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INTRODUCTION
The Animal Investigation Program (AIP) developed an operational plan to
support the activities proposed for the Supplemental Test Site (STS).
This plan involved the collection of tissue samples from two mule deer
and three beef cattle for radioanalysis and histopathologic studies prior
to Project Faultless. Each event was to be followed by the collection of
a like number of animals and tissue samples if there was a likelihood
that an exposure to radioactive isotopes had occurred. Collection of
animals and techniques of radioanalysis were to be the same as those
employed in past studies.* Tissues submitted for histopathology included
heart, kidneys, liver, lung, striated muscle, spleen, thyroid, and any
other tissue appearing grossly abnormal. A thin section was taken of
each organ and preserved in formalin prior to submission to Dr. Gordon
Solomon at Colorado State University for microscopic examination.
Just prior to the Faultless Event approximately 5,000 sheep were grazing
10 to 25 miles from Ground Zero. A U. S. Public Health Service (USPHS)
certified monitor and the AIP veterinarian were stationed with the sheep-
herders prior to and during the event to advise the sheepherders of the
time of the event and document any damages that might occur from ground
shock or venting. Since there was no venting and only moderate ground
shock involved, no animals were collected following the Faultless Event.
*Southwestern Radiological Health Laboratory Intralaboratory Technical
Report ITR-11.
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PROCEDURES
Arrangements were made with Mr. Grube, owner of the K Ranches, to pur-
chase three head of cattle grazing in the STS area prior to the scheduled
event. These cattle were then transported to the Nevada Test Site (NTS),
Well #3, USPHS facilities for sacrifice.
As an item of general interest, it should be pointed out that animals
purchased for analysis are frequently culls that are of questionable
value to the owner due to existing gross pathological processes.* Even
though gross pathology is often present, market price is paid for these
animals in order to maintain good public relations with the livestock
producers in the area.
Additionally, permission was obtained from the Nevada Fish and Game com-
mission to collect two mule deer from the Morey Bench Area (Figure 1).
This area was selected because of its proximity to the test area and also
because it is the winter habitat of large herds of deer.
These animals were shot and dissected according to an established protocol
and subjected to post-mortem examinations. Tissue samples were collected
for radioanalysis and histopathological examination as outlined in
SWRHL ITR-11. Gamma spectroscopy and/or radiochemistry of the various
biological samples included analysis for 131I, 1Q6Ru5 144ce, 54Mn, 140Ba,
137cs, 65Zn, 89sr, 90sr, 95zr, and total potassium.**
Tissues collected for radioanalysis were abomasal content, abomasal tissue,
proximal 6" of the femur, liver, lung, muscle, rumen contents, thyroid,
and hock joint.
Tissues collected for histopathology were heart, kidneys, liver, lung,
muscle, spleen, thyroid, and any tissues which appeared grossly abnormal.
*Cancer eye (squamous cell carcinoma) and physical deformities.
**Potassium-40 is found in physiologically normal tissues at a fixed per-
cent (0.0118) with stable potassium (K). Within this report, potassium
values are calculated from the measured radioactive potassium using the
0.0118 ratio and are recorded as total potassium.
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Proposed Test Site
Morey Bench
Figure 1. Proposed Supplemental Test Site and Morey Bench.
3
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RADIQANALYSIS RESULTS AND DISCUSSION
Radioanalysis of the tissues from the mule deer and the cattle revealed
very low or nondetectable levels of all radionuclides except for stron-
tium-89 and -90 in the tarsal and femoral bones of the respective species
and 1311 in the thyroids of the deer (Appendix A). The greatest variety
of radionuclides occurred in the rumen and abomasum contents (Table A-3
and A-4). Rumen contents yielded 131l in the deer, 144ce in two cows
and the deer, 14°Ba in the deer, l°6Ru in one deer, 65zn in one cow,
and 95Zr in one deer. Abomasal content yielded 131l in two cows, 144ce
in a cow and a deer, 140Ba in a cow, l°6Ru in two cows and a deer, l37Cs
in a cow, 65Zn in a cow, and 95zr -jn two cows and a deer. The abomasal
tissue contained 54^n and 65Zn in one cow and l°6Ru in another cow
(Table A-5). The liver contained 54Mn and 95zr in one cow and !37Cs in
a deer (Table A-6). Muscle contained ^Ce and 54Mn in one cow each and
137Cs, 65Zn, and 95Zr in a deer (Table A-2). There were no radionuclides
present in any of the lungs analyzed (Table A-7). Femoral bone samples
from the cows and tarsal bones from the deer--all revealed the presence
of strontium-89 and -90 (Table A-8). With the exception of 131l, radio-
nuclides values were equal to or below those reported in tissues col-
lected during routine slaughters of cattle from the Knoll Creek, Delamar
Valley, and NTS areas and deer collected on the NTS. The source of the
131I found in the deer thyroids is thought to have come from atmospheric
fallout resulting from non-United States nuclear tests conducted on
24 December 1967.*
The levels of radioactive isotopes reported in these animals have little
significance other than baselines for comparison with animals collected
following events conducted on the Supplemental Test Site. Should there
be a significant delay prior to further testing, additional animals should
be collected to determine if any change has occurred in the levels of
isotopes reported from this sampling especially in view of the higher
than expected 131l levels.
*Southwestern Radiological Health Laboratory Air Surveillance Network,
January 1968 Air Results.
4
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PATHOLOGY RESULTS AND DISCUSSION
Pathologic lesions occurring in these animals were consistent with pre-
viously reported lesions found during other routine slaughters (BER-8 and
BER-18). The observed lesions (see Appendix B) can be attributed to
advanced age, natural disease processes, or traumatic injury.
Lesions in the deer were limited to sarcosporidiosis in skeletal and
cardiac muscles and focal atelectasis in the lungs. The only gross
lesion* observed at the time of necropsy was pleural adhesions to the
right lobe of one lung.
Significant gross lesions in the cattle included ecchymotic hemorrhages
in the lungs, fibrous adhesions between the liver and diaphragm, gall
stones, subcutaneous botfly larvae, and a cystic kidney. Microscopic
lesions included sarcosporidiosis in cardiac and skeletal muscles;
pyelonephritis and several foci of lymphocytic infiltration suggestive
of lymphosarcoma in one kidney; microcysts, fibrosis, and mineralization
of collecting tubules in another kidney; foci of lymphocytic accumula-
tions and focal atelectasis in the lungs; focal hypertrophy and hyper-
plasia of reticuloendothelial cells in the liver; and squamous cell
carcinoma of the cornea! epithelium.
*With the exception of gunshot damage.
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SUMMARY
Animals collected on the STS were subjected to thorough radioanalytic
and histopathologic examinations. Results indicate that these animals
are comparable to animals collected during other routine sacrifices--
both in pathologic lesions present and detectable body burdens of
selected radioisotopes--except for the increased level of ^lj -jn the
thyroids of the deer and one of the cows. A tentative explanation of
this exception could possible be a result of fallout from Non-United
States nuclear tests. Presence of the other radioisotopes was erratic
and only slightly above background levels. The pathologic lesions
observed were attributable to degenerative and/or acquired conditions
commonly seen with advanced age, traumatic injury, metabolic disturb-
ances, or infection.
Since (as expected) there was no release of radioactivity from the
Faultless event, no animals were collected following the event for
post-mortem analysis.
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APPENDIX
Appendix A Radioanalysis Results — Bovine and Mule Deer
Appendix B Necropsy Protocols
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APPENDIX A
RADIOANALYSIS RESULTS-BOVINE AND MULE DEER
Table A~l. Thyroid.
144Ce 54Mn 140Ba 106Ru 137Cs 65Zn 95Zr K
AIP NO. _ pCi/g pCi/g pCi/g pCi/g pCi/g pCi/g pCi/g pCi/g g/g
BOV-1-N14-68
BOV-2-N14-68
BOV-3-N14-68
MD-1-N14-68
MD-2-N14-68
ND
8.45
ND
148
204
ND
ND
ND
ND
ND
1.36
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND = Nondetectable
Table A-2. Muscle.
AIP NO.
BOV-1-N14-68
BOV-2-N14-68
BOV-3-N14-68
MD-1-N14-68
MD-2-N14-68
pCi/g
ND
ND
ND
ND
ND
l44Ce
pCi/g
ND
ND
0.043
ND
ND
54Mn
pCi/g
ND
0.051
ND
ND
ND
l40Ba
pCi/g
ND
ND
ND
ND
ND
106Ru
pCi/g
ND
ND
ND
ND
ND
13/Cs
pCi/g
ND
ND
ND
ND
0.092
6bZn
pCi/g
ND
ND
ND
ND
1.84
9bZr
pCi/g
ND
ND
ND
ND
0.065
K
g/g
0.0048
0.0031
0.0035
ND
0.0032
ND = Nondetectable
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Table A-3. Rumen Contents.
AIP NO.
BOV-1-N14-68
BOV-2-N14-68
BOV-3-N14-68
MD-1-N14-68
MB-2-N14-68
pci/g
ND
ND
ND
0.101
0.100
144Ce
pCi/g
0.585
0.065
ND
0.904
1.990
b4Mn
pCi/g
ND
ND
ND
ND
ND
140Ba
pCi/g
ND
ND
ND
0.071
0.111
iUbRu
pCi/g
ND
ND
ND
ND
0.246
pCi/g
ND
ND
ND
ND
ND
pCi/g
ND
2.59
ND
ND
ND
95 Zr
pCi/g
ND
ND
ND
ND
1.12
K
g/g
ND
0.0010
ND
0.0019
ND
ND = Nondetectable
Table A-4. Abomasum
AIP NO.
BOV-1-N15-68
BOV-2-N14-68
BOV-3-N14-68
MD-1-N14-68
MD-2-N14-68
pCi/g
0.071
0.209
ND
ND
ND
Contents.
144Ce
pci/g
ND
1.35
ND
2.25
ND
54Mn
pCi/g
ND
ND
ND
ND
ND
140Ba
pCi/g
ND
0.095
ND
ND
ND
106Ru
pCi/g
ND
0.221
0.105
0.484
ND
137Cs
pCi/g
ND
0.224
ND
ND
ND
65Zn
pCi/g
ND
1.95
ND
ND
ND
95Zr
pCi/g
0.149
0.097
ND
1.89
ND
K
g/g
0.0014
0.0135
ND
0.0029
ND
ND = Nondetectable
Table A- 5. Abomasum
AIP NO.
BOV-1-N14-68
BOV-2-N14-68
BOV-3-N14-68
MD-1-N14-68
MD-2-N14-68
pCi/g
ND
ND
ND
ND
ND
Tissue,
144Ce
pCi/g
ND
ND
ND
ND
ND
b4Mn
pCi/g
ND
0.057
ND
ND
ND
14UBa
pCi/g
ND
ND
ND
ND
ND
106Ru
pCi/g
ND
ND
0.154
ND
ND
137Cs
pCi/g
ND
ND
ND
ND
ND
65Zn
pCi/g
ND
0.081
ND
ND
ND
95Zr
pCi/g
ND
ND
ND
ND
ND
K
g/g
ND
0.0015
ND
ND
ND
ND = Nondetectable
8
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Table A-6. Liver.
AIP NO. pCi/g
BOV-1-N14-68 ND
BOV-2-N14-68 ND
BOV-3-N14-68 ND
MD-l^N14-68 ND
MD-2-N14-68 ND
ND = Nondetectable
Table A-7. Lung.
AIP NO. pCi/g
BOV-1-N14-68 NA
BOV-2-N14-68 NA
BOV-3-N14-68 ND
MD-1-N14-68 ND
MD-2-N14-68 ND
NA = Not Analyzed
ND = Nondetectable
Table A-8. Bone.
AIP NO.
BOV-1-N14-68
BOV-2-N14-68
BOV-3-N14-68
MD-1-N14-68
MD-2-N14-68
144^ 54
Ce Mn
pCi/g pCi/g
ND 0.048
ND
ND
ND
ND
144
Ce
pCi/g
NA
NA
ND
ND
ND
89Sr
3.672
2.173
3.783
2.461
2.899
ND
ND
ND
ND
54Mn
Pci/g
NA
NA
ND
ND
ND
pCi/g
± 0.
± 1.
± 1.
± 1.
± 1.
pCi/g pCi/g
ND ND
ND ND
ND ND
ND ND
ND ND
140Ba 106Ru
pCi/g pCi/g
NA NA
NA NA
ND ND
ND ND
ND ND
Bone Ash
853*
275
197
064
648
I37rc 657n
Cs Zn
pCi/g pCi/g
ND ND
ND ND
ND ND
0.058 ND
ND ND
137Cs 65Zn
pCi/g pCi/g
NA NA
NA NA
ND ND
ND ND
ND ND
90Sr pCi/g
5.809 ± 0.
14.532 ± 0.
15.473 ± 0.
9.877 ± 0.
23.217 ± 0.
pCi/g
0.049
ND
ND
ND
ND
9bZr
pCi/g
NA
NA
ND
ND
ND
Bone
461*
715
710
625
993
K
g/g
0.0026
0.0023
0.0023
0.0029
ND
K
g/g
NA
NA
0.0024
ND
ND
Ash
* Counting error
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APPENDIX B
NECROPSY PROTOCOLS
1. AIP No. BOV-1-N14-68
a. History. This animal was born and spent its entire life on the
Site "C" Supplemental Test Site open range area.
DATE OF DEATH: 10 Jan. 1968 HOUR: 0900 LOCATION: Well 3, NTS
DATE OF NECROPSY: 10 Jan. 1968 HOUR: 0930 LOCATION: Well 3, NTS
SEX: Male AGE: 1 year
SPECIES: Bovine BREED: Hereford
b. External Appearance. This is a long yearling Hereford steer in
poor physical condition. Both hind feet are deformed at their distal
ends. The horny covering of the third and fourth digits of the left
hind foot are missing and have been replaced by a fibrous pad. The
horny coverings of the third and fourth digits of the right hind foot
have a convex shape. Due to this abnormal shape, the animal walked
on the volar surface of the sole near the area of the digital cushion
resulting in a heavy fibrous covering in this area.
c. Respiratory System. Diffuse ecchymotic hemorrhages are seen in
the right and left apical and cardiac lobes.
d. Miscellaneous. Hypoderma bovis larvae are found in the sub-
dermal tissue of the back.
e. Necropsy Results.
(1) Liver: There is hypertrophy and focal hyperplasia of the
reticuloendothelial cells.
(2) Kidney: Focal accumulation of lymphocytes interstitially
which is suggestive of early lymphosarcoma.
10
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2. Alf No. BOV-2-N14-68
a. History. This animal was born and spent its entire life on the
Site "C" Supplemental Test Site open range area.
DATE OF DEATH: 10 Jan. 1968 HOUR: 0900 LOCATION: Well 3, NTS
DATE OF NECROPSY: 10 Jan. 1968 HOUR: 1015 LOCATION: Well 3, NTS
SEX: Female AGE: 13 years
SPECIES: Bovine BREED: Hereford
b. External Appearance. This is a mature, nonlactating Hereford
cow in poor physical condition. Carcinomas are seen on the cornea
of both eyes.
c. Digestive System. There are extensive fibrous adhesions between
the peritoneal surface of the liver and the serosa of the diaphragm.
Approximately 10 small stones, 5 mm in diameter, are seen in the
gall bladder.
d. Genitourinary System. The uterus contains a fetus which is in
approximately the third month of gestation.
e. Necropsy Results.
(1) Cardiac Muscle: Moderate sarcosporidiosis is present.
(2) Kidney: Multiple foci of accumulated immature and mature
lymphocytes suggestive of lymphosarcoma. There are focal
areas of proximal pyelonephritis characterized by the
presence of polymorphonuclear leukocytes in the tubules.
(3) Striated Muscle: There is slight sarcosporidiosis.
(4) Liver: There is focal hypertrophy and hyperplasia of
reticuloendothelial cells.
(5) Eye: Squamous cell carcinoma which originates from the
corneal epithelium.
11
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3. AIP No. BOV-3-N14-68
a. History. This animal was born and spent its entire life on the
Site "C" Supplemental Test Site open range area.
DATE OF DEATH: 10 Jan. 1968 HOUR: 0900 LOCATION: Well 3, NTS
DATE OF NECROPSY: 10 Jan. 1968 HOUR: 1100 LOCATION: Well 3, NTS
SEX: Female AGE: 14 years
SPECIES: Bovine BREED: Hereford
b. External Appearance. This is a mature Hereford cow in poor
physical condition.
c. Digestive System. Extensive fibrous adhesions are seen between
the peritoneal surface of the diaphragm and the serosa of the liver.
d. Genitourinary System. The kidney contains two small fluid-filled
cysts approximately 3 mm in diameter. Tissues of these areas are
submitted for histopathologic study.
e. Necropsy Results.
(1) Striated Muscle: There is moderate sarcosporidiosis.
(2) Cardiac Muscle: There is moderate sarcosporidiosis.
(3) Kidney: There are several foci of fibrosis or chronic
nephritis. In addition, there are microcysts characterized
by the absence of kidney parenchyma and the crowding to
together of peripheral structures. There is also mineral-
ization of many collecting tubules.
(4) Lung: There are foci of lymphocytic accumulations sub-
pi eurally and within septa.
12
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4. AIP No. MD-1-N14-68
a. History. This animal was collected in the Morey Bench area.
DATE OF DEATH: 16 Jan. 1968 HOUR: 2130
LOCATION: 4-Mile Canyon (STS)
DATE OF NECROPSY: 16 Jan. 1968 HOUR: 2130
LOCATION: 4-Mile Canyon (STS)
SEX: Female AGE: 4 years WEIGHT: 125 Ib. METHOD: Estimation
SPECIES: Odocoileus heminus COMMON NAME: Mule Deer
b. External Appearance. This is a female deer in good condition.
The left humerus is fractured (due to bullet).
c. Genitourinary System. The uterus contains two fetuses estimated
to be in the first trimester of gestation.
d. Respiratory System. There are diffuse ecchymotic hemorrhages
throughout all lobes of the lung. The left apical lobe is macerated
by a bullet.
e. Necropsy Results.
(1) Lung: Extensive alveolar hemorrhage and free blood within
the bronchioles.
(2) Skeletal Muscle: Slight sarcosporidiosis.
5. AIP No. MD-2-N14-68
a. History. This animal was collected in the Morey Bench area.
DATE OF DEATH: 17 Jan. 1968 HOUR: 2000
LOCATION: 4-Mile Canyon (STS)
13
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DATE OF NECROPSY: 17 Jan. 1968 HOUR: 2000
LOCATION: 4-Mile Canyon (STS)
SEX: Female AGE: 2 1/2 years WEIGHT: 100 Ib.
METHOD: Estimation
SPECIES: Odocoileus hemionus COMMON NAME: Mule Deer
b. External Appearance. This is a female deer in good condition.
c. Genitourinary System. The uterus contains a fetus estimated
to be in the first trimester of gestation.
d. Respiratory System. All lobes of the lung contain diffuse
ecchymotic hemorrhages. The left apical lobe is completely
macerated by the bullet. The right lobes of the lung have fibrous
adhesions extending from their peripheral border to the chest wall
e. Necropsy Results.
(1) Lung: There are focal areas of atelectasis.
(2) Cardiac Muscle: There is slight sarcosporidiosis.
14
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DISTRIBUTION
1 - 20 SWRHL, Las Vegas, Nevada
21 James E. Reeves, Manager, NVOO/AEC, Las Vegas, Nevada
22 Robert H. Thalgott, NVOO/AEC, Las Vegas, Nevada
23 Chief, NOB, DASA, NVOO/AEC, Las Vegas, Nevada
24 Donald H. Edwards, Effects Safety Division, NVOO/AEC,
Las Vegas, Nevada
25 R. C. Emens, NTS Support Office, NVOO/AEC, Mercury, Nevada
26 Martin B. Biles, DOS, USAEC, Washington, D. C.
27 John S. Kelly, DPNE, USAEC, Washington, D. C.
28 Philip Allen, ESSA/ARL, NVOO/AEC, Las Vegas, Nevada
29 G. D. Ferber, ARL/ESSA, Silver Spring, Maryland
30 - 34 Charles L. Weaver, NCRH, PHS, Rockville, Maryland
35 Bernd Kahn, NCRH, RATSEC, Cincinnati, Ohio
36 Northeastern Radiological Health Lab., Winchester, Mass.
37 Southeastern Radiological Health Lab. Montgomery, Alabama
38 Donald W. Hendricks, Safety Evaluation Div., NVOO/AEC,
Las Vegas, Nevada
39 Mail & Records, NVOO/AEC, Las Vegas, Nevada
40 Paul T. Tueller, University of Nevada, Reno, Nevada
41 F. Ward Whicker, Colorado State University, Fort Collins, Colo.
42 V. R. Bohman, University of Nevada, Reno, Nevada
43 Al Jonez, U. S. Bureau of Reclamation, Boulder City, Nevada
44 District Ranger, U. S. Forest Service, Las Vegas, Nevada
45 Region Supervisor, Nevada Fish & Game Commission, Las Vegas, Nev.
46 Director, Nevada Fish & Game Commission, Reno, Nevada
47 R. S. Davidson, Battelle Memorial Institute, Columbus, Ohio
48 - 49 DTIE, U. S. AEC, Oak Ridge, Tennessee
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