Impacts of radiation exposure on the bacterial and fungal microbiome of small mammals in the Chernobyl Exclusion Zone.

Environmental impacts of the 1986 Chernobyl Nuclear Power Plant accident are much debated, but the effects of radiation on host microbiomes have received little attention to date. We present the first analysis of small mammal gut microbiomes from the Chernobyl Exclusion Zone in relation to total absorbed dose rate, including both caecum and faeces samples. We provide novel evidence that host species determines fungal community composition, and that associations between microbiome (both bacterial and fungal) communities and radiation exposure vary between host species. Using ambient versus total weighted absorbed dose rates in analyses produced different results, with the latter more robust for interpreting microbiome changes at the individual level. We found considerable variation between results for faecal and gut samples of bank voles, suggesting faecal samples are not an accurate indicator of gut composition. Associations between radiation exposure and microbiome composition of gut samples were not robust against geographical variation, although we identified families of bacteria (Lachnospiraceae and Muribaculaceae) and fungi (Steccherinaceae and Strophariaceae) in the guts of bank voles that may serve as biomarkers of radiation exposure. Further studies considering a range of small mammal species are needed to establish the robustness of these potential biomarkers.

Analysis of 129I and 127I in soils of the Chernobyl Exclusion Zone, 29 years after the deposition of 129I.

The Chernobyl Exclusion Zone (CEZ) represents a unique natural laboratory that received significant 129I contamination across a range of soils and land-use types in a short time period in 1986. Data are presented on 129I and 127I in soil samples collected from highly contaminated areas in the CEZ in 2015. The geometric mean (GM) total concentration of stable iodine (127I) was 6.7 × 10-7 g g-1 and the (GM) total concentration of 129I was 2.39 × 10-13 g g-1, equivalent to 1.56 mBq kg-1. GM total 127I concentration is below the European average soil concentration of 3.94 × 10-6 g g-1, while 129I is significantly higher than the pre-Chernobyl activity concentration for 129I of 0.094 mBq kg-1. Significant differences were found in the extractability of native, stable 127I and 129I almost 30 years after the introduction of 129I to the soils. Both 127I and 129I were predominantly associated with alkaline-extractable soil organic matter, established using a three-step sequential extraction procedure. Whereas 127I was significantly correlated with gross soil organic matter (measured by loss on ignition), however, 129I was not. The ratio of 129I/127I was significantly lower in extracts of soil organic matter than in more labile (soluble and adsorbed) fractions, indicating incomplete equilibration of 129I with native 127I in soil humic substances after 29 years residence time in the CEZ soils. The initial physico-chemical form of 129I in the CEZ soils is unknown, but the widespread presence of uranium oxide fuel particles is unlikely to have influenced the environmental behaviour of 129I. Our findings have implications for long-term radiation dose from 129I in contaminated soils and the use of native, stable 127I as a proxy for the long-term fate of 129I.

Genome-wide DNA methylation changes in two Brassicaceae species sampled alongside a radiation gradient in Chernobyl and Fukushima.

The long-term radiological impact to the environment of the nuclear accidents in Chernobyl and Fukushima is still under discussion. In the course of spring of 2016 we sampled two Brassicacea plants, Arabidopsis thaliana and Capsella bursa-pastoris native to Ukraine and Japan, respectively, alongside a gradient of radiation within the exclusion and difficult to return zones of Chernobyl (CEZ) and Fukushima (FEZ). Ambient dose rates were similar for both sampling gradients ranging from 0.5 to 80 µGy/h at plant height. The hypothesis was tested whether a history of several generations of plants growing in enhanced radiation exposure conditions would have led to changes in genome-wide DNA methylation. However, no differences were found in the global percentage of 5-methylated cytosines in Capsella bursa pastoris plants sampled in FEZ. On the other hand a significant decrease in whole genome methylation percentage in Arabidopsis thaliana plants was found in CEZ mainly governed by the highest exposed plants. These data support a link between exposure to changed environmental conditions and changes genome methylation. In addition to methylation the activity concentration of different radionuclides, 137Cs, 90Sr, 241Am and Pu-238,239,240 for CEZ and 137, 134Cs for FEZ, was analysed in both soil and plant samples. The ratio of 5.6 between 137Cs compared to 134Cs was as expected five years after the FEZ accident. For CEZ 137Cs is the most abundant polluting radionuclide in soil followed by 90Sr. Whereas 241Am and Pu-isotopes are only marginally present. In the plant tissue, however, higher levels of Sr than Cs were retrieved due to a high uptake of 90Sr in the plants. The 90Sr transfer factors ranged in CEZ from 5 to 20 (kg/kg) depending on the locality. Based on the activity concentrations of the different radionuclides the ERICA tool was used to estimate the total dose rates to the plants. It was found that for FEZ the doses was mainly contributable to the external Cs-isotopes and as such estimated total dose rates (0.13-38 µGy/h) were in the same range as the ambient measured dose rates. In strong contrast this was not true for CEZ where the total dose rate was mainly due to high uptake of the 90Sr leading to dose rates ranging from 1 to 370 µGy/h. Hence our data clearly indicate that not taking into account the internal contamination in CEZ will lead to considerable underestimation of the doses to the plants. Additionally they show that it is hard to compare the two nuclear accidental sites and one of the main reasons is the difference in contamination profile.

Elevated mitochondrial genome variation after 50 generations of radiation exposure in a wild rodent.

Currently, the effects of chronic, continuous low dose environmental irradiation on the mitochondrial genome of resident small mammals are unknown. Using the bank vole (Myodes glareolus) as a model system, we tested the hypothesis that approximately 50 generations of exposure to the Chernobyl environment has significantly altered genetic diversity of the mitochondrial genome. Using deep sequencing, we compared mitochondrial genomes from 131 individuals from reference sites with radioactive contamination comparable to that present in northern Ukraine before the 26 April 1986 meltdown, to populations where substantial fallout was deposited following the nuclear accident. Population genetic variables revealed significant differences among populations from contaminated and uncontaminated localities. Therefore, we rejected the null hypothesis of no significant genetic effect from 50 generations of exposure to the environment created by the Chernobyl meltdown. Samples from contaminated localities exhibited significantly higher numbers of haplotypes and polymorphic loci, elevated genetic diversity, and a significantly higher average number of substitutions per site across mitochondrial gene regions. Observed genetic variation was dominated by synonymous mutations, which may indicate a history of purify selection against nonsynonymous or insertion/deletion mutations. These significant differences were not attributable to sample size artifacts. The observed increase in mitochondrial genomic diversity in voles from radioactive sites is consistent with the possibility that chronic, continuous irradiation resulting from the Chernobyl disaster has produced an accelerated mutation rate in this species over the last 25 years. Our results, being the first to demonstrate this phenomenon in a wild mammalian species, are important for understanding genetic consequences of exposure to low-dose radiation sources.

Effects of radionuclide contamination on leaf litter decomposition in the Chernobyl exclusion zone.

The effects of radioactive contamination on ecosystem processes such as litter decomposition remain largely unknown. Because radionuclides accumulated in soil and plant biomass can be harmful for organisms, the functioning of ecosystems may be altered by radioactive contamination. Here, we tested the hypothesis that decomposition is impaired by increasing levels of radioactivity in the environment by exposing uncontaminated leaf litter from silver birch and black alder at (i) eleven distant forest sites differing in ambient radiation levels (0.22-15µGyh(-1)) and (ii) along a short distance gradient of radioactive contamination (1.2-29µGyh(-1)) within a single forest in the Chernobyl exclusion zone. In addition to measuring ambient external dose rates, we estimated the average total dose rates (ATDRs) absorbed by decomposers for an accurate estimate of dose-induced ecological consequences of radioactive pollution. Taking into account potential confounding factors (soil pH, moisture, texture, and organic carbon content), the results from the eleven distant forest sites, and from the single forest, showed increased litter mass loss with increasing ATDRs from 0.3 to 150µGyh(-1). This unexpected result may be due to (i) overcompensation of decomposer organisms exposed to radionuclides leading to a higher decomposer abundance (hormetic effect), and/or (ii) from preferred feeding by decomposers on the uncontaminated leaf litter used for our experiment compared to locally produced, contaminated leaf litter. Our data indicate that radio-contamination of forest ecosystems over more than two decades does not necessarily have detrimental effects on organic matter decay. However, further studies are needed to unravel the underlying mechanisms of the results reported here, in order to draw firmer conclusions on how radio-contamination affects decomposition and associated ecosystem processes.

Radioadaptive response following in utero low-dose irradiation.

Acute radiation exposure is known to cause biological damage that leads to severe health effects. However, the effects and subsequent health implications of exposure to low doses of ionizing radiation are unclear. The purpose of this study was to investigate the effects of low-dose ionizing radiation exposures in utero. Pregnant laboratory mice (BALB/c) were exposed to low-dose Chernobyl radiation [10-13 mSv per day for 10 days] during organogenesis. The progeny were born and weaned in an uncontaminated laboratory, then were exposed to an acute radiation dose (2.4 Sv). Analysis of our end points (litter dynamics, DNA damage, bone marrow stem cell function, white blood cell counts and gene expression) suggests that a low-dose (100-130 mSv) in utero exposure to ionizing radiation is not deleterious to the offspring. Rather DNA damage, white blood cell levels, and gene expression results suggest a radioadaptive response was elicited for the in utero exposure with respect to the effects of the subsequent acute radiation exposure.

Overview of the cooperation between the Chernobyl Center's International Radioecology Laboratory in Slavutych, Ukraine, and U.S. research centers between 2000 and 2010.

The International Radioecology Laboratory (IRL) located in Slavutych, Ukraine, was created in 1999 under the initiative of the United States Government and the Government of Ukraine in the framework of international cooperation on evaluation and minimization of consequences of the Chernobyl nuclear power plant (ChNPP) accident. Since the time the IRL was founded, it has participated in a large number of projects, including the following: 1) study of radionuclide accumulation, distribution, and migration in components of various ecological systems of the Chernobyl Exclusion Zone (ChEZ); 2) radiation dose assessments; 3) study of the effects of radiation influence on biological systems; 4) expert analysis of isotopic and quantitative composition of radioactive contaminants; 5) development of new methods and technologies intended for radioecological research; 6) evaluation of future developments and pathways for potential remediation of the ChEZ areas; 7) assistance in provision of physical protection systems for ionizing irradiation sources at Ukrainian enterprises; 8) reviews of open Russian language publications on issues associated with consequences of the ChNPP accident, radioactive waste management, radioecological monitoring, and ChNPP decommissioning; 9) conduct of training courses on problems of radioecology, radiation safety, radioecological characterization of test sites and environmental media, and research methods; 10) conduct of on-site scientific conferences and workshops on the ChEZ and radioecology problems; participation in off-site scientific conferences and meetings; and 11) preparation of scientific and popular science publications and interactions with mass media representatives. This article provides a brief overview of the major achievements resulting from this cooperation between the IRL and U.S. research centers.

Radiation dose assessment for the biota of terrestrial ecosystems in the shoreline zone of the Chernobyl nuclear power plant cooling pond.

Radiation exposure of the biota in the shoreline area of the Chernobyl Nuclear Power Plant Cooling Pond was assessed to evaluate radiological consequences from the decommissioning of the Cooling Pond. This paper addresses studies of radioactive contamination of the terrestrial faunal complex and radionuclide concentration ratios in bodies of small birds, small mammals, amphibians, and reptiles living in the area. The data were used to calculate doses to biota using the ERICA Tool software. Doses from 90Sr and 137Cs were calculated using the default parameters of the ERICA Tool and were shown to be consistent with biota doses calculated from the field data. However, the ERICA dose calculations for plutonium isotopes were much higher (2-5 times for small mammals and 10-14 times for birds) than the doses calculated using the experimental data. Currently, the total doses for the terrestrial biota do not exceed maximum recommended levels. However, if the Cooling Pond is allowed to draw down naturally and the contaminants of the bottom sediments are exposed and enter the biological cycle, the calculated doses to biota may exceed the maximum recommended values. The study is important in establishing the current exposure conditions such that a baseline exists from which changes can be documented following the lowering of the reservoir water. Additionally, the study provided useful radioecological data on biota concentration ratios for some species that are poorly represented in the literature.

Method for simultaneous 90Sr and 137Cs in-vivo measurements of small animals and other environmental media developed for the conditions of the Chernobyl exclusion zone.

To perform in vivo simultaneous measurements of the 90Sr and 137Cs content in the bodies of animals living in the Chernobyl Exclusion Zone (ChEZ), an appropriate method and equipment were developed and installed in a mobile gamma beta spectrometry laboratory. This technique was designed for animals of relatively small sizes (up to 50 g). The 90Sr content is measured by a beta spectrometer with a 0.1-mm-thick scintillation plastic detector. The spectrum processing takes into account the fact that the measured object is "thick-layered" and contains a comparable quantity of 137Cs, which is a characteristic condition of the ChEZ. The 137Cs content is measured by a NaI scintillation detector that is part of the combined gamma beta spectrometry system. For environmental research performed in the ChEZ, the advantages of this method and equipment (rapid measurements, capability to measure live animals directly in their habitat, and the capability of simultaneous 90Sr and 137Cs measurements) far outweigh the existing limitations (considerations must be made for background radiation and the animal size, skeletal shape, and body mass). The accuracy of these in vivo measurements is shown to be consistent with standard spectrometric and radiochemical methods. Apart from the in vivo measurements, the proposed methodology, after a very simple upgrade that is also described in this paper, works even more accurately with samples of other media, such as soil and plants.

Frequency distributions of 90Sr and 137Cs concentrations in an ecosystem of the "Red Forest" area in the Chernobyl exclusion zone.

In the most highly contaminated region of the Chernobyl Exclusion Zone, the "Red Forest" site, the accumulation of the major dose-affecting radionuclides (90Sr and 137Cs) within the components of an ecological system encompassing 3,000 m(2) was characterized. The sampled components included soils (top 0-10 cm depth), Molina caerulea (blue moor grass), Camponotus vagus (carpenter ants), and Pelobates fuscus (spade-footed toad). In a comparison among the components of this ecosystem, the 90Sr and 137Cs concentrations measured in 40 separate grids exhibited significant differences, while the frequency distribution of the values was close to a logarithmically-normal leptokurtic distribution with a significant right-side skew. While it is important to identify localized areas of high contamination or "hot spots," including these values in the arithmetic mean may overestimate the exposure risk. In component sample sets that exhibited logarithmically normal distribution, the geometric mean more accurately characterizes a site. Ideally, risk assessment is most confidently achieved when the arithmetic and geometric means are most similar, meaning the distribution approaches normal. Through bioaccumulation, the highest concentrations of 90Sr and 137Cs were measured in the blue moor grass and spade-footed toad. These components also possessed distribution parameters that shifted toward a normal distribution.

Radiation ecology issues associated with murine rodents and shrews in the Chernobyl exclusion zone.

This article describes major studies performed by the Chernobyl Center's International Radioecology Laboratory (Slavutich, Ukraine) on radioecology of murine rodents and shrews inhabiting the Chernobyl Exclusion Zone. The article addresses the long-term (1986-2005) and seasonal dynamics of radioactive contamination of animals and reviews interspecies differences in radionuclide accumulations and factors affecting the radionuclide accumulations. It is shown that bioavailability of radionuclides in the "soil-to-plant" chain and a trophic specialization of animals play key roles in determining their actual contamination levels. The total absorbed dose rates in small mammals significantly reduced during the years following the Chernobyl Nuclear Power Plant accident. In 1986, the absorbed dose rate reached 1.3-6.0 Gy h(-1) in the central areas of the Chernobyl Exclusion Zone (the "Red Forest"). In 1988 and 1990, the total absorbed dose rates were 1.3 and 0.42 Gy h(-1), respectively. In 1995, 2000, and 2005, according to the present study, the total absorbed dose rates rarely exceeded 0.00023, 0.00018, and 0.00015 Gy h(-1), respectively. Contributions of individual radiation sources into the total absorbed dose are described.

Environmental radiation monitoring in the Chernobyl exclusion zone--history and results 25 years after.

This paper describes results of the radiation environmental monitoring performed in the Chernobyl Exclusion Zone (ChEZ) during the period following the 1986 Chernobyl Nuclear Power Plant accident. This article presents a brief overview of five comprehensive reports generated under Contract No. DE-AC09-96SR18500 (Washington Savannah River Company LLC, Subcontract No. AC55559N, SOW No. ON8778) and summarizes characteristics of the ChEZ and its post-accident status. The history of development of the radiation monitoring research in the ChEZ is described also. This paper addresses the characteristics of radiation monitoring in the ChEZ, its major goals and objectives, and changes in these goals and objectives in the course of time, depending on the tasks associated with the phase of mitigation of the ChNPP accident consequences. The results of the radiation monitoring in the ChEZ during the last 25 years are also provided.

Assessment of (90)sr and (137)cs penetration into reinforced concrete (extent of "deepening") under natural atmospheric conditions.

When assessing the feasibility of remediation following the detonation of a radiological dispersion device or improvised nuclear device in a large city, several issues should be considered, including the levels and characteristics of the radioactive contamination, the availability of resources required for decontamination and the planned future use of the city's structures and buildings. Currently, little is known about radionuclide penetration into construction materials in an urban environment. Knowledge in this area would be useful when considering costs of a thorough decontamination of buildings, artificial structures and roads in an affected urban environment. Pripyat, a city substantially contaminated by the Chernobyl Nuclear Power Plant accident in April 1986, may provide some answers. The main objective of this study was to assess the depth of (90)Sr and (137)Cs penetration into reinforced concrete structures in a highly contaminated urban environment under natural weather conditions. Thirteen reinforced concrete core samples were obtained from external surfaces of a contaminated building in Pripyat. The concrete cores were drilled to obtain sample layers of 0-5, 5-10, 10-15, 15-20, 20-30, 30-40 and 40-50 mm. Both (90)Sr and (137)Cs were detected in the entire 0-50 mm profile of the reinforced cores sampled. In most of the cores, over 90% of the total (137)Cs inventory and 70% of the total (90)Sr inventory was found in the first 0-5 mm layer of the reinforced concrete. Strontium-90 ((90)Sr) had penetrated markedly deeper into the reinforced concrete structures than (137)Cs.

The effects of environmental low-dose irradiation on tolerance to chemotherapeutic agents.

The nuclear disaster at Chernobyl, Ukraine, in April of 1986 continues to impact the environment on many different levels. Studies of epidemiological, environmental, and genetic impacts have been prolific since the accident, revealing interesting results concerning the effects of radiation. The long-tailed field mouse, Apodemus flavicollis, was collected from distinct localities near the Chernobyl site and evaluated based on in vivo responses to the current clinically employed chemotherapeutic agents bleomycin (BLM) and vinblastine (VBL), as well as the immune modulator lipopolysaccharide (LPS). Maximum tolerable doses of three different cancer drugs were administered to the rodents from three different lifestyles: native mice living and reproducing in a radioactive environment, native mice living and reproducing in an uncontaminated region, and laboratory-reared mice (Mus musculus BALB/c) with a known sensitivity to the chemical agents tested. The endpoints employed include micronucleus formation, immune cell induction, differential gene expression, and chemotherapeutic side effects such as lethargy and weight loss. In accordance with the well-studied phenomenon termed radio-adaptation, we observed varied tolerance to chemotherapeutic treatment dependent on history of ionizing radiation exposure. The results of the present study demonstrate a differential response to chemotherapeutic treatment with respect to previous levels of radiation exposure, suggesting a potential benefit associated with low-dose radiation exposure. Data reported herein could have a profound impact on the development of novel cancer treatments involving low-dose ionizing radiation.

Understanding the genetic consequences of environmental toxicant exposure: Chernobyl as a model system.

We sampled vole populations in Ukraine with the dual goal of characterizing population diversity and of providing a biogeographic perspective to evaluate experimental designs used for previous studies. Our data indicate that genetic diversity in bank vole populations is widely variable across regions and that diversity estimates in contaminated sites are unremarkable compared to those in uncontaminated areas. Furthermore, the relative frequencies of haplotypes have remained statistically identical throughout multiple sampling periods. Thus, the genetic data from bank vole populations in Ukraine fail to support the hypothesis that mutational changes in contaminated regions are the product of exposure to Chernobyl radiation. Our results suggest that genetic diversity in radioactive regions of Ukraine is probably a function of natural geographic variation rather than increased mutational pressure from radiation exposure and underscore the importance of adequate geographic sampling in studies designed to elucidate the effects of toxicant exposure.

Variation in mitochondrial DNA control region haplotypes in populations of the bank vole, Clethrionomys glareolus, living in the Chernobyl environment, Ukraine.

Bank vole, Clethrionomys glareolus, specimens have been annually sampled from the radioactive Chernobyl, Ukraine, environment and nonradioactive reference sites since 1997. Exposed voles continually exhibit increased mitochondrial DNA haplotype (h) and nucleotide diversity (ND), observed in the hypervariable control region (1997-1999). Increased maternal mutation rates, source-sink relationships, or both are proposed as hypotheses for these differences. Samples from additional years (2000 and 2001) have been incorporated into this temporal study. To evaluate the hypothesis that an increased mutation rate is associated with increased h, DNA sequences were examined in a phylogenetic context for novel substitutions not observed in haplotypes from bank voles from outside Ukraine or in other species of Clethrionomys. Such novel substitutions might result from in situ mutation events and, if largely restricted to samples from radioactive environments, support an increased maternal mutation rate in these areas. The only unique substitution meeting this criterion was found in an uncontaminated reference site. All other substitutions are found in other haplotypes of the bank vole or in other species. Increased maternal mutation rates do not appear to explain trends in h and ND observed in northern Ukraine. Studies examining ecological dynamics will clarify the reasons behind, and significance of, increased levels of h in contaminated areas.

Predicting radionuclide transfer to wild animals: an application of a proposed environmental impact assessment framework to the Chernobyl exclusion zone.

A number of assessment frameworks have been proposed to provide a mechanism to demonstrate protection of the environment from ionising radiation. Whilst some of these are being used for assessment purposes they have largely not been validated against field measurements. In this paper we compare the predictions of transfer parameters recommended by one of these frameworks (FASSET) with observed whole-body 90Sr and radiocaesium activity concentrations in a range of mammal and invertebrate species sampled within the Chernobyl exclusion zone. Predicted activity concentrations were generally within the observed ranges and mean predictions for reference organisms were similar to, or circa one order of magnitude higher than, the observed means. However, some predictions were more than one order of magnitude lower than observed values. No data were available to test predictions for the other radionuclides released by the Chernobyl accident. In a separate paper the outputs of this assessment will be used to estimate doses to reference organisms and compare these to observed radiation induced effects reported within the Chernobyl zone.

Exposure to chronic, low-dose rate gamma-radiation at Chornobyl does not induce point mutations in Big Blue mice.

Empirical genetic effects resulting from low-dose rate irradiation and chronic, cumulative exposure are poorly characterized. Expected effects are based on epidemiological studies and downward, linear extrapolations from nonthreshold models derived from acute, high-dose exposures. These extrapolations and their associated risk coefficients have no experimental support, and because of their inherent uncertainty they are the subject of considerable debate. The expectation of deleterious genetic effects resulting from low-dose rate irradiation and chronic exposure is in need of empirical assessment because this type of exposure is typical of those encountered in occupational, residential, and environmental settings. Recent acute low-dose (<10 cGy) studies using cytogenetic and point mutation endpoints indicate that observed effects range from those lower than spontaneous to an increase in the frequency of point mutations. Using the Big Blue assay, we examined the ability of chronic, continuous gamma-irradiation (2.3 x 10(-3) cGy/min) in the Chornobyl environment to induce point mutations. This system has demonstrated a significant point mutation sensitivity (4.5-fold increase) to acute, high-dose (1-3 Gy) gamma-radiation. Mutant frequencies and the mutation spectra were examined in exposed and reference samples of Big Blue mice following 90 days exposure (cumulative absorbed dose = 3 Gy) to the Chornobyl environment. No significant increase in the mutant frequency or bias in the mutational spectrum was observed in exposed individuals. This finding suggests that low-dose rate gamma-irradiation at Chornobyl does not induce point mutations and that cumulative, chronically absorbed doses do not induce the same genetic effects as acute doses of the same magnitude.

Mitochondrial DNA heteroplasmy in laboratory mice experimentally enclosed in the radioactive Chernobyl environment.

Mitochondrial DNA heteroplasmy using the protein-coding cytochrome b (Mtcyb) gene was assessed in laboratory mice (C57BL/6 and BALB/c) exposed to the Chernobyl environment. Subacute to subchronic (30-40 days) exposure resulted in a cumulative radiation dose of 1.2-1.6 Gy ( approximately 0.04 Gy/day). Mice were sampled prior to introduction into the enclosures and again after removal from the enclosures. Nucleotide variation (site heteroplasmy) in 306 pre-exposure Mtcyb gene copies (122400 base pairs) was compared to variation in 354 postexposure gene copies (141600 base pairs). Five mutant copies, each characterized by a single nucleotide substitution, were observed (four in the pre-exposure samples, one in a postexposure sample). The frequencies of mutant gene copies and nucleotide substitutions in pre-exposure and postexposure samples were not significantly different. This suggests that this type of exposure (i.e. low dose rate) does not pose a significant mutation risk to the Mtcyb gene in digit tissue. Furthermore, no significant radiation risk to analogous human tissues may exist when occupational exposures involve low dose rates such as these. Finally, linear, cumulative models of genetic risk currently used to estimate radiation-induced effects are likely to be inappropriate for low-dose-rate exposures and need to be re-evaluated critically.