Effects of ionising radiation exposure on plants, fish and mammals: relevant data for environmental radiation protection.

In order to develop a framework for the assessment of the environmental impact of radiation, it is necessary to establish the relationship between exposure (dose rate, accumulated dose) and the effects that may be induced in plants and animals. With this purpose in mind, the data available on effects induced by ionising radiation in various wildlife groups have been reviewed as part of the FASSET project. This paper has highlighted that the available information on the effects of low dose rate, continuous irradiation (< 10(3) microGy h(-1)) is reasonable for plants, fish and mammals, but is scarce or non-existent for other wildlife groups. Thus, the effects induced in plants, fish and mammals after chronic exposure to radiation are presented in this paper. The fragmentary nature of the available, relevant information has made it very difficult to characterise the desired dose rate-response relationships in any detail. However, it can be broadly concluded that, although minor effects may be seen at lower dose rates in the most sensitive species and systems, the threshold for statistically significant effects in most studies is about 10(2) microGy h(-1). The responses then increase progressively with increasing dose rate and usually become very clear at dose rates > 10(3) microGy h(-1) sustained for a large fraction of the lifespan.

DNA damage in zebrafish larvae induced by exposure to low-dose rate gamma-radiation: detection by the alkaline comet assay.

This study has determined the sensitivity of the alkaline comet assay for the detection of strand breaks in the DNA of cells taken from a whole organism rather than a single cell type as in previously reported studies. The assay has been performed on cells from whole zebrafish larvae irradiated for 1 or 24 h at dose rates of 0.4, 1.2 or 7.2 mGy/h. Zebrafish larvae exposed to only 1.2 mGy/h of gamma-radiation for 1h showed a statistically significant increase in DNA damage compared to controls. This represents a high sensitivity of this animal model for DNA damage and of the comet assay protocol used for detecting such damage. Increasing the exposure time from 1 to 24 h caused significant increases in DNA damage in zebrafish larvae, although the modest size of these increases in damage for the relatively large increases (24 times) in total absorbed dose indicates that dose rate may be the major factor in determining the level of DNA damage observed under the conditions of these experiments.

Long-term irradiation of a marine fish, the plaice Pleuronectes platessa: an assessment of the effects on size and composition of the testes and of possible genotoxic changes in peripheral erythrocytes.

PURPOSE: Previous investigation showed the very significant effects of chronic gamma-radiation on plaice testes at mean absorbed dose rates as low as 1.3 mGy h(-1) given over a period of 168 days (accumulated dose 4.7 Gy). The present paper examines the effects on the testes of exposure to even lower dose rates of gamma-radiation given over periods of 73 days or 197 days. In addition, the use of micronucleus counts and flow-cytometric measurement of nuclear DNA content in samples of peripheral blood for monitoring genotoxic effects has been assessed. MATERIALS AND METHODS: In Experiment 1, adult male plaice were exposed at mean absorbed dose rates of 0.25, 0.5 or 1.2 mGy h(-1) for 73 days (mean accumulated doses of 0.43, 0.85 and 2.03 Gy, respectively) and in Experiment 2 to 0.24, 0.5 or 1.0 mGy h(-1) for 197 days (mean accumulated doses of 1.07, 2.24 and 4.57 Gy, respectively). At termination the testes were removed, weighed and sections were prepared and examined histometrically. In addition, in Experiment 2, blood samples were taken during exposure and at termination. Blood smears were scored for micronuclei and samples processed and examined for nuclear DNA content by flow cytometry. RESULTS: Significant reductions in testis weight were seen in all radiation groups after 197 days of exposure, which were predominantly due to decreased amounts of sperm. In plaice killed after 73 days (at an earlier stage of spermatogenesis), there were no significant differences in weight compared with controls but amounts of spermatogonia were significantly reduced in irradiated fish. CONCLUSIONS: Exposure to dose rates as low as 0.24 mGy h(-1) of gamma-radiation given over a period of 197 days significantly reduced the weights of plaice testes, this being consequent on reductions in the amounts of sperm. Although there was some evidence of radiation affecting the numbers of spermatogonia it was not possible to determine the primary target for radiation damage which eventually caused the sperm reductions. Along with the related work described by Greenwood and Knowles (1996) this is the first investigation of a marine fish and it indicates that plaice testes are probably more radiosensitive than those previously described in tropical fish and of a similar radiosensitivity to mammalian testes. Although significant effects were observed after the lowest dose rate used of 0.24 mGy h(-1), this is still a factor of about 400 times greater than the estimated absorbed dose rate to plaice testes in the north-east Irish Sea off Sellafield at the present time. Micronucleus counts and flow-cytometric analysis of blood DNA both failed to show any evidence of genotoxic damage.

Effect of chronic irradiation on the humoral immune response of a marine fish, the eelpout (Zoarces viviparus L.).

Eelpout (Zoarces viviparus), a marine fish, were exposed to gamma-radiation at a dose-rate of 2.0 mGy h-1. They were injected with the antigen trinitrophenyl-keyhole limpet haemocyanin (TNP-KLH) and their specific humoral immune response to the TNP hapten measured. There were no differences in the primary antibody response between unirradiated controls and treated fish. However, the response to a second injection of antigen was significantly increased in irradiated fish. There were no significant radiation effects on total serum protein or serum lysozyme. Radiation effects on testis were also examined and there was a significant radiation-induced reduction in relative weight (gonadosomatic index). These results are discussed with reference to environmental radiation levels in areas of authorized radioactive waste disposal.

Functional changes in the pig kidney following irradiation with fractionated doses of fast neutrons (42 MeVd-->Be).

The right kidney of female Large White pigs, approximately 14 weeks old, was irradiated with fractionated doses of fast neutrons (42 MeVd-->Be). The total doses used were 6.6-9.2 Gy. Changes in kidney function, assessed as the functional index (FI, where FI = irradiated kidney function/unirradiated kidney function) or as individual kidney glomerular filtration rate (GFR) and effective renal plasma flow (ERPF), were serially determined up to 104 weeks after irradiation using 99Tcm-DTPA and 131I-hippuran renography. The animals were then euthanized, the kidneys removed and weighed. A dose-dependent reduction in FI was seen within 13 weeks of irradiation. Measuring individual kidney function revealed a hyperaemic response in both irradiated and unirradiated kidney 4 weeks after irradiation. This was followed by a dose-dependent reduction in irradiated kidney GFR and particularly ERPF. The ED50 value for the impairment in ERPF, assessed as the percentage of irradiated kidneys exhibiting a > or = 50% reduction in ERPF, was significantly lower than that for GFR, i.e. 7.20 +/- 0.10 Gy compared with 8.44 +/- 0.07 Gy (p < 0.001). A dose-related reduction in irradiated kidney weight was also observed. These fast neutron-induced changes in renal function and weight are qualitatively similar to those observed following photon irradiation of the pig kidney.

The effect of chronic radiation on the humoral immune response of rainbow trout (Onchorhynchus mykiss Walbaum).

Two separate experiments have examined the effect of exposing rainbow trout to chronic gamma-radiation, commencing immediately after fertilization. In experiment 1 the period of exposure extended for 20 days with groups receiving mean dose rates of 1.87, 3.73 and 9.03 mGy h-1, and mean total accumulated doses of 0.83, 1.66 and 4.01 Gy respectively. At 5 months of age fish were tested for specific antibody response to dinitrophenol coupled to keyhole limpet haemocyanin (DNP-KLH) and there was no significant difference in titre between irradiated groups and unirradiated controls. In experiment 2 the exposure period was extended to 246 days from fertilization. Mean dose-rates to the three groups used were the same as in the first experiment until hatching at 21 days and then lower with rates of 0.99, 1.9, and 4.66 mGy h-1 to the free-swimming fish. The mean total accumulated doses over the whole irradiation period were 5.43, 10.53 and 25.43 Gy respectively. The antibody response to DNP-KLH was significantly lower in trout receiving the highest dose-rate when compared with those of unirradiated controls or the lowest dose-rate group. The significance of these results is discussed in relation to radiation levels in areas of radioactive waste disposal, and results from a similar study published previously.

The relative biological effectiveness of fractionated doses of fast neutrons (42 MeVd----Be) for normal tissues in the pig. IV. Effects on renal function.

The effects of fractionated doses of fast neutrons (42 MeVd----Be) on the radiation response of the pig kidney have been assessed and compared with those observed after X irradiation. Following X irradiation there was a marked increase in the total dose at which renal function was preserved with decreasing fraction size. The rate of this increase was dependent on the overall treatment time; for fractionated irradiation given over 18 or 39 days the exponents related to fraction number, N, were 0.36 +/- 0.03 and 0.48 +/- 0.003, respectively. In contrast, there was no significant change in the iso-effect dose for renal injury following fractionated irradiation with fast neutrons where there was also little effect of varying the overall treatment time. Analysing these data by means of the linear-quadratic (LQ) model, using both an Fe-plot and the Tucker test, gave alpha/beta ratios of 2.42 +/- 0.06 Gy and 2.99 +/- 0.16 Gy, respectively, for X-ray doses given in 18 days. For fractionated doses of X rays given in 39 days the alpha/beta ratios were 0.40 +/- 0.01 Gy and 0.47 +/- 0.02 Gy, respectively. The alpha/beta ratios for renal tissue following fast neutron irradiation obtained by the two methods were also similar, i.e. 15.00 +/- 0.60 Gy and 15.72 +/- 3.76 Gy, respectively. The pronounced fractionation effect seen with X irradiation, particularly for doses administered over 39 days as opposed to 18 days, coupled with the absence of any such effect with fast neutrons, resulted in a marked increase in relative biological effectiveness (RBE) with decreasing X-ray dose/fraction. The slopes of the resulting regression lines were -0.73 +/- 0.05 and -0.33 +/- 0.02, respectively. The lack of dose sparing associated with fractionation, or variation of the overall treatment time for fast neutron irradiation, suggests that doses administered to tumours adjacent to the kidney can be given as a few relatively large dose/fractions in a short overall treatment time without an increased risk of complications related to renal tissue. This may be of therapeutic advantage in the treatment of rapidly proliferating tumours where dose may be wasted using more conventional protracted fractionated irradiation schedules.

The relative biological effectiveness of fractionated doses of fast neutrons (42 MeVd----Be) for normal tissues in the pig. I. Effects on the epidermis and dermal vascular/connective tissues.

The effects of fractionated doses of fast neutrons (42 MeVd----Be) on the early epithelial and later dermal response of pig skin have been assessed and compared with those after X irradiation. For the early epithelial reaction, i.e. moist desquamation, the relative biological effectiveness (RBE) of the neutron beam increased with the decreasing size of the X-ray dose/fraction. There was an experimentally observed upper RBE value of approximately 2.75 for X-ray doses/fraction of between 2 and 5 Gy. For the late reaction of ischaemic dermal necrosis the RBE was greater than 3.0 for X-ray doses/fraction of less than 3 Gy and, based on the assumptions made in the linearquadratic model of cell survival, an upper limiting RBE of 4.32 +/- 0.39 was calculated for infinitely small doses/fraction. These findings were compared with other radiobiological data and the conclusions drawn from the results of clinical trials. It was concluded that for the sparing of late effects in skin and subcutaneous tissues, relative to acute reactions, a relatively small number of fractions in a short overall treatment time may be optimal for fast neutron therapy.

Experimental irradiation of the rat ureter: the effects of field size and the presence of contrast medium on incidence and latency of hydronephrosis.

Following X-irradiation of a 1.5 cm length of rat ureter, hydronephrosis developed after doses down to 10 Gy. The estimated ED50 was 11.8 Gy. In the dose range 37.4 Gy to 17.5 Gy there was a significant increase in latency with decreasing dose, but at lower doses the latency did not increase further. Reducing the length of ureter irradiated to 0.5 cm or 0.8 cm caused a decrease in incidence of hydronephrosis and longer latency periods. The ED50 for rats irradiated to 0.5 cm of ureter was 29.6 Gy. The possibility that secondary radiation produced as a result of interaction between X-radiation and iodinated contrast medium might affect the radiation induction of hydronephrosis was investigated. No difference was found between groups of rats irradiated with or without injection of contrast media.

Radiation-induced hydronephrosis in the rat: a new experimental model.

An experimental model for investigating the effects of localized X-irradiation of a single ureter or the bladder trigone in rats is described. Obstruction of the urinary tract in the irradiated region gives rise to hydroureter and hydronephrosis and the development of these, as detected urographically, gives a clear-cut end point. After irradiation of the ureter with a single dose of 37.4 Gy many rats died of gut lesions but after 23.4 Gy only one such death occurred while 14 of 16 rats developed hydronephrosis. Irradiation of the bladder trigone was not associated with intercurrent deaths, even after 40 Gy, and after 25 Gy 9 of 11 rats developed hydronephrosis.

Changing sensitivity of neonatal rats to tumorigenic effects of N-nitroso-N-ethylurea and X-radiation, given singly or combined.

After neonatal injection of rats with 10 mg N-nitroso-N-ethylurea [(ENU) CAS: 759-73-9]/kg body weight, whole-body X-irradiation with 1.25 Gy X-radiation caused a reduction in induced neurogenic tumors which was dependent on the interval between the two treatments. The reduction was greatest when radiation was given 1 day after ENU and progressively decreased with irradiation at 5 and 30 days. Changes in radiosensitivity in the neonatal period were also seen for ovarian tumors and squamous cell carcinomas of the mouth. Ovarian tumors were induced by 1.25 Gy X-rays alone, with a higher incidence occurring after irradiation at 5 days than at 30 days of age. Squamous cell carcinomas of the mouth have a high spontaneous incidence in this strain of rat, but 1.25 Gy X-radiation caused a significant reduction which was greater after irradiation at 5 than at 30 days of age. Pituitary tumors occurred in untreated and treated rats with a higher incidence in females. Only in males was there any suggestion of an effect of the various treatments on pituitary tumor incidence.

Bone in the irradiated lung of the guinea-pig.

Considerable amounts of metaplastic bone developed in the lungs, and to a lesser extent, the liver of guinea-pigs which had received X-radiation to these organs. The bone was lamellar and had bone marrow cavities containing haemopoietic tissue.

Reduction of N-nitroso-N-ethylurea-induced neurogenic tumors by X-radiation: a life-span study in rats.

Whole-body X-irradiation after neonatal injection with N-nitroso-N-ethylurea (ENU) significantly reduced the incidence of induced neurogenic tumors in inbred HMT rats kept for their complete life-span. After administration of 10 mg ENU/kg and 1.25 Gy X-radiation, the incidence of schwannomas but not of gliomas was reduced as compared to the incidence in rats given 10 mg ENU/kg only. In contrast, after administration of 4 mg ENU/kg, 1.25 Gy reduced the incidence of gliomas but not of schwannomas. Administration of 1.25 Gy alone induced a remarkably high incidence of rats with neurogenic tumors (20%). Latency of tumor detection was not significantly affected by radiation. Among the most frequently occurring nonneurogenic tumors, squamous cell carcinomas were reduced in incidence by treatment with ENU, 1.25 Gy X-radiation, or both combined. No treatment affected the incidence of pituitary or mammary tumors. There was a preponderance of ovarian tumors in rats given 4 mg ENU/kg + 1.25 Gy. An incidental finding was the occurrence of granular cell tumors in 7 rats from different treatment groups.

The radiosensitivity of the guinea-pig spinal cord to X-rays: the effect of retreatment at one year and the effect of age at the time of irradiation.

Day-old guinea-pigs were given a non-paralysing dose of 10 Gy X-rays to the lumbar spinal cord. One year later there was no evidence of any residual radiation damage as the dose required to produce paralysis was the same for these animals as for others not previously irradiated. When given a single dose only, guinea-pigs irradiated when 1 day old became paralysed after lower doses and with shorter latencies than those irradiated at 1 year. When irradiated at 30 days of age, the dose inducing paralysis was the same as at 1 year of age, but the latency period was shorter and similar to guinea-pigs irradiated at 1 day old. Thus at 30 days of age, adult radiotolerance had been acquired but latencies were still as short as in neonates. Whatever the age at irradiation, changes in latency for paralysis were closely related to changes in histopathological lesions in the cord and both were related to dose. White matter necrosis always occurred after higher doses and after shorter latencies than diffuse vacuolar demyelination.

The relative biological effectiveness of fast neutrons (42MeVd leads to Be) for early and late normal tissue injury in the pig.

Early and late radiation damage has been investigated in a number of normal tissues in the pig after irradiation with single doses of neutrons produced by 42MeV deuterons on beryllium. The results have been compared with data obtained after irradiation with single doses of 250kV X rays. In the skin a low RBE value of approximately 1.2 was obtained for the early (3-9 week) epithelial reaction. For the subsequent dermal vascular response, higher RBE values in the range of 1.35-1.6 were obtained: the RBE decreasing with an increase in the neutron dose. For late skin damage, assessed by the relative reduction in the linear dimensions of an irradiated field, a RBE value of approximately 1.5 was obtained. In the kidney the RBE value, for a neutron dose level (550 cGy) at which renal function was just preserved, was 2.0. A lower value of 1.7 was found for doses resulting in a loss of renal function. The results of 133Xenon clearance studies showed two waves of impaired ventilation function in the irradiated lung. In the acute reaction (3-9 months), at a dose level consistent with just preserving normal ventilation function, the RBE value was less than 1.2. For late lung damage (15-24 months) the RBE value was higher, 1.4. For the rectum, methods are presently only available for assessing acute damage. A RBE of 2.0 was found for neutron doses in the range 350-575 cGy. The RBE values for early endpoints in the skin, lung and gut of the pig are comparable with those published previously for other species, including man. The values for late effects in pig skin and lung were higher than for early damage in those tissues.

The effect of x-radiation given after neonatal administration of ethyl nitrosourea on incidence of induced nervous system tumours.

Neonatal rats were injected with neurocarcinogenic amounts of ethyl nitrosourea (ENU) and whole-body X-irradiated 24 h later. An absorbed radiation dose of 1.25 Gy caused a statistically significant decrease in the incidence of nervous system tumours induced by ENU (10 mg/kg). Absorbed doses of 0.2 Gy or 1.25 Gy given after ENU (4 mg/kg) also reduced their incidence, but not significantly so. The X-irradiation did not affect the range of histological appearances amongst the tumours but malignant schwannomas, particularly those of the trigeminal nerve, were significantly reduced by 1.25 Gy given after ENU (10 mg/kg). The mean latency for clinical signs of tumour appearance was not affected by radiation. A small number of nervous system tumours occurred in rats given neonatal X-radiation only and it seems highly likely that these were radiation-induced.