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 ------- 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 ------- 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. ------- 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 ------- 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 ------- 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. ------- 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. ------- Proposed Test Site Morey Bench Figure 1. Proposed Supplemental Test Site and Morey Bench. 3 ------- 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 ------- 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. ------- 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. ------- APPENDIX Appendix A Radioanalysis Results — Bovine and Mule Deer Appendix B Necropsy Protocols ------- 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 ------- 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 ------- 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 ------- 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 ------- 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 ------- 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 ------- 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 ------- 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 ------- 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 ------- |