An Overview of Research in Radiation Biology and Protective Measures in the Manhattan Project During the War Years 1942-1946

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AN OVERVIEW OF RESEARCH IN RADIATION BIOLOGY AEJD PROTECTIVE MEASURES I N THE MANHATTAN PROJECT DURING THE WAR YEARS. IV.UNIVERSITY OF WASHINGTON PROJECT

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UNIVERSITY OF WASHINGTON PROJECT .Within a few weeks after the decision to establish the Hanford Engineer Works in western Washington was made, Col. Warren and Gen. Groves were considering plans to study fish in the Columbia and Snake rivers in the state of Washington. A number of reasons for these studies were readily apparent. One, the vast fisheries program should in no way be endangered. TWO, even though the prospect of small doses of radioactivity suggested that no identifiable changes in fish might be expected, it was nevertheless considered important to have records of studies on the response of fish to various doses of radiation.A third reason was that an additional biological specimen could be pursued to develop the general understanding of radiation biology and enhance the studies already underway in other areas of the Manhattan Project. In order to maintain secrecy and to avoid connection with the Corps of Engineers,the Manhattan District Division of Medicine arranged to have the contract which was eventually to be consumated with the University of Washington, negotiated through the Office of Scientific Research and Development (OSRD).In this way the relationship between the secret nuclear development a t Hanford and the University of Washington might be obscured fora number of years.Accordingly, Lauren Donaldson was

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invited to Washington to meet with Col. Warren and a representative from OSRD in the person of Harry T. Wenzel, technical aide to the executive committee of OSRD, which a t that time was headed by James B. Conant. The studies on salmon could conceivably have been carried out at another site,but because of the prior intensive interest in the fisheries program at the University of Washington and because Gen. Groves had at one time been a student there,the studies were naturally directed to that location. The University and Dr.Donaldson duly established the Applied Fisheries Laboratory and x - ray equipment was installed shortly thereafter.It is of interest that the installation of the x - ray equipment was supervised by a member of the University of Rochester. The studies were initiated promptly after the I t was made clear to Dr.Donaldson that the primary objective was to study the effect of radiation on fish,butin order t o obscure the fact that intensive studies in radiation biology were to be pursued, The program was titled, " Investigation of - the Use of X-Rays in the Treatment of Fungoid Infections in the Most of the work was performed at the University of Washington,

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but because of considerable interest in,and the importance of, studying the rivers themselves a small aquatic laboratory was opened at Hanford under Richard Foster ' s direction. facility studies on the concentration of radioactivity in the water and studies of plankton were pursued, among others. An outline of the work is given in the following pages.It is a concise compilation of the nature of the project,the data collected and their early evaluation. This work was initiated in August,1943,and continued to be the major effort of the laboratory during the war. Many irradiated fish were released and the major tasks left to be completed in the future were the recovery of the marked fish from irradiated parents and the determination of the success of The following is a general outline of the protocols and the nature of the experiments. A. Adult Chinook Salmon from the Green River (Females and Males)4 - Spawning occurred during October and November of 1943. - Deaths occurred during October and November of1943 and were

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natural in males only ; females were sacrificed when - Total number of eggs 55,084 - Release and Descending Migration.A total of 4,835 (100 r > finger ling were released in the Samish River on May 11, 1944, and a total of 4,844 control finger ling were released there on the following day. It was originally planned to release Inthe summer of 1946, the commercial fishery yielded2 chinook salmon bearing the marks of the experiment.In addition to these two saltwater re ' cover ies, 23 marked adults returned to the Samish River in the fall of 1946. Of these, 12 (including only2females) were controls and the remaining 11 (also including only2females)were B. Adult Sockeye Salmon from Cult us Lake, B.C. (Females & Males) - 24 fish were treated in Novemebr, 1943; there were 10 controls. - Spawning occurred during November and December of 1943. * The system of marking for identification utilized in these experiments involved the removal of the adipose and the left ventral fin of treated fish and the removal of the adipose and right ventral fin of control fish.

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- Deaths occurred during November and December of 1943 and were natural for males only ; females were sacrificed when finger ling were released in Cult us Lake on February22 and 27, 1945and a total of 4,139 control finger ling were released there on February20and22,1945.Returns were c. Chinook Salmon Eggs from the Green River containing931to 934 eggs. Two lots were allowed for each exposure and2lots were controls. D - Chinook Salmon Eyed Eggs from the Green River Two lots were allowed for each

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E.Silver Salmon from the Green River (Males Only) - One fish was treated at each exposure in January, 1944; there were2controls for each exposure. - Deaths occurred during January and February of 1944 and were natural. The effects on skin and certain other tissues were also studied. - 5,400 finger ling were treated i n May of 1944, with600 in each lot. - Tissue for sectioning obtained on a weekly basis. - Statistical analyses were made of the effects upon mortality, weight,length,cell counts in circulating blood and he ma top oie tic cell counts in the kidney.Tissue samples were preserved at the time of the experiment for use in further his to logical studies. G . Rainbow Trout Yearlings from the University of Washington each lot, except the1,500rlot(16)and the 2,500 rlot (121,

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with18tagged controls and 2 untagged ones which were spawned atthe same time. - Studies on mortality,growth and pathological conditions were carried out at the same time. Most of the fish were still alive in 1946. _- The trout spawned fora second time in the spring of 1946 and the offspring were saved.Adults were planted in Green Lake inthe summer of 1946. - First spawn from the rainbow trout yearlings totaled115,401 eggs. The eggs were secured from 6 7 of the71 females and divided into 134 lots. Growth abnormalities in spawning t o eyed eggs and morat lit y were studied.Feeding was continued. H. Adult Chinook Salmon from the Columbia River (Females & Males) - Spawning occurred in September, 1944. - Deaths were natural.Thirty - three fish were obtained from Spring Creek and transferred 270 miles by truck which greatly reduced the vitality of the adults and their progeny.Thirty - three percent of the mortality was a result of transfer and - bl or tal it y,growth and weight data were collected. - Total number of eggs 54,347

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I. Adult Steelhead Trout from the Columbia River (Leavenworth) - Spawning occurred during May and June,1944. - Deaths occurred during May and June, 1944 and were natural for both females and males. - Total number of eggs 50,107 of the control group 3,285 were marked. The 25rand 50 r groups were not marked before releasing. - Release and Descending Migration.A total of 13,360 unmarked fish were planted in the Leavenworth Hatchery in September, 1944. The marked fish were transferred to Leavenworth and liberated in the No.3holding pond i n The summary of results was essentially recorded in the following Adult Fish.No observable change in any species was noted after exposures to25, 50 or 100 r. A definite increase in

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mortality of trout exposed to1000ror more was found. very little distinct evidence of damage in terms of growth or weight change was shown. Some cyto logical changes of a transient nature were observed in the gut and the anterior kidney (base of the he ma top oie tic systems).In small goldfish, the changes occurred at about the same level(100r)with Eggs. I n the eyed stage (in which eye pigmentation first begins to show) there was a definite retardation in growth a t 1000 r, although his to logical change was first apparent at a lower level. (250r).As in the above, the most sensitive tissues were the gut and the anterior kidney.A50%mortality figure occurred at approximately the1000r level.At a level in excess of this,but almost certainly less than the radiation dose of2500r used in the protocol, 100% mortality was reached. Finger ling s.Retardation in growth became apparent at500r with his to logical changes appearing at this level in the spleen and the anterior kidney.In this stage,the spleen and anterior kidney were sites of blood formation.At1250r approximately 25%mortality was reached for the several species.At some point in excess of 2500 r, a100%mortality was reached. * A modest study of the immediate response of goldfish to radiation was also done.

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Unfortunately, no level of an intermediate stage between 2500 r and 500 r was used. In the radiation of parent fish at less than 500 r, no readily identifiable changes had been observed in the number spawned, fertility,abn or mail ty of developing eggs, etc. Significant changes were not observed up to the1000r level as noted Statistical analyses of all data collected in the above studies was carried out. It was intended that data be collected throughout1948 on the results of the fish that had gone outto sea and would eventually return to spawn,. Chemical and Temperature Changes Both fishes and eggs exposed to the effluent unrefrigerated, refrigerated and in a three - t o - one concentration showed a n increase in mortality of a significant degree. Some of the chemicals also increased mortality due to actual chemical injury to gill structures and possible respiration by the egg. An increase in temper tau redid not influence the severity of changes, but hastened their onset. After living in the effluent(pile) water for as long as two months under controlled conditions,the absorbed and deposited

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radioactivity was measured.In no instance was the level great enough to produce identifiable damage. The major portion of the deposition was in the form of short - lived products. Only traces of the long - lived radioactive materials were found. As correlated with the effect of external radiation,the dosage received was negligible compared t o that required to produce identifiable damage. It is useful to insert here some data from a recent publication : were exposed to various levels of continuous gamma irradiation to determine the level at which the return of these adults as individuals would be affected. Large numbers of eggs(63,000 to140,000)were used for each treatment,along with an equal number of controls. The dose rate ranged from 0.5r / day in each of the later years.After rearing for about90days,the control and irradiated groups were differently marked and released for their migration to salt water. On their return 2to6yr later,length,weight and sex data were determined on each adult salmon. No significant change was noted in adult returns until radiation levels of 10r / day and above were used.At these