High-Accuracy Relative Biological Effectiveness Values Following Low-Dose Thermal Neutron Exposures Support Bimodal Quality Factor Response with Neutron Energy.

Theoretical evaluations indicate the radiation weighting factor for thermal neutrons differs from the current International Commission on Radiological Protection (ICRP) recommended value of 2.5, which has radiation protection implications for high-energy radiotherapy, inside spacecraft, on the lunar or Martian surface, and in nuclear reactor workplaces. We examined the relative biological effectiveness (RBE) of DNA damage generated by thermal neutrons compared to gamma radiation. Whole blood was irradiated by 64 meV thermal neutrons from the National Research Universal reactor. DNA damage and erroneous DNA double-strand break repair was evaluated by dicentric chromosome assay (DCA) and cytokinesis-block micronucleus (CBMN) assay with low doses ranging 6-85 mGy. Linear dose responses were observed. Significant DNA aberration clustering was found indicative of high ionizing density radiation. When the dose contribution of both the 14N(n,p)14C and 1H(n,γ)2H capture reactions were considered, the DCA and the CBMN assays generated similar maximum RBE values of 11.3 ± 1.6 and 9.0 ± 1.1, respectively. Consequently, thermal neutron RBE is approximately four times higher than the current ICRP radiation weighting factor value of 2.5. This lends support to bimodal peaks in the quality factor for RBE neutron energy response, underlining the importance of radiological protection against thermal neutron exposures.

Assessing effects of legacy nuclear waste on plants: Sensitive fern (Onoclea sensibilis) gametophyte viability at the Chalk River site.

We evaluated the effects of chronic exposure to environmental radiological contamination on the reproductive fitness of sensitive fern (Onoclea sensibilis) by quantifying viability in haploid gametophytes of spores collected from ferns from background and contaminated areas of the Chalk River site. Dose rates measured in situ at field sites ranged from 60 to 849 µGy h-1, with effects possible at the more contaminated sites (greater than 400 µGy h-1). Fern spores were also irradiated from 1 to 1000 Gy to develop dose-response curves. We found no effects on gametophyte viability at the most contaminated areas of the Chalk River site, where we estimated growing season doses of 0.3-3.7 Gy. Dose-response curves show evidence of hormesis, with an increase in gametophyte viability up to 10 Gy, followed by a rapid decline to no viable gametophytes at doses of 1000 Gy. The sensitive fern is not a radiosensitive plant species, but effects do occur within the normal range (10-1000 Gy) of most plant species, making it useful as a sentinel species from a community perspective. Sensitive fern spore germination is high and stable over field dose ranges, with effects primarily on gametophyte viability. This method shows promise as an effects monitoring tool for sites with radiological contamination.

Sr-90 soil to plant transfer factor reduction using calcium and polymer soil amendments.

Introducing calcium into soils can inhibit Sr-90 uptake by plants. To test the efficacy of calcium amendments on the inhibition of Sr-90 uptake by edible plants, a number of different calcium applications, including calcium nitrate, calcium thiosulfate and a mixture of both liquid solutions, were used in this study. Pea plants (Pisum sativum 'Sabre') grown in Sr-90 contaminated soil from seeds to maturity were watered with these calcium solutions. Two different polymers, one inert and one nutrient enriched, were incorporated into the contaminated soil where pea seeds were sowed to ascertain a continuous supply of calcium and essential nutrients. Results show that the heterogeneity of Sr-90 distribution in soil translated to disparate Sr-90 contents in plant tissues. However, on average, irrigation with calcium solutions in conjunction with the usage of polymers consistently yielded a reduction in Sr-90 uptake by the plants. The lowest soil-to-plant transfer factor (TF) values were measured in the edible pea part of the plant, followed by the flowers, roots, stems, pea shells and then leaves. TF values for pea shells were between 4.9 and 20.9, and between 0.3 and 2.8 for the peas. Results do not allow the identification of one particular chemical solution that would systematically be the best choice to minimize Sr-90 uptake.

Effects of in vivo exposure to tritium: a multi-biomarker approach using the fathead minnow, Pimephales promelas.

Tritium (3H) is a radioactive isotope of hydrogen. In the environment, the most common form of tritium is tritiated water (HTO). However, tritium can also be incorporated into organic molecules, forming organically bound tritium (OBT). The present study characterized the effects of tritium on the health of the fathead minnow, Pimephales promelas. Fish were exposed to a gradient of HTO (activity concentrations of 12,000, 25,000, and 180,000 Bq/L) and OBT using food spiked with tritiated amino acids (OBT only, with an activity concentration of 27,000 Bq/L). A combined exposure condition where fish were placed in 25,000 Bq/L water and received OBT through feed was also studied. Fish were exposed for 60 days, followed by a 60-day depuration period. A battery of health biomarkers were measured in fish tissues at seven time points throughout the 120 days required to complete the exposure and depuration phases. HTO and OBT were also measured in fish tissues at the same time points. Results showed effects of increasing tritium activity concentrations in water after 60 days of exposure. The internal dose rates of tritium, estimated from the tissue free-water tritium (TFWT) and OBT activity concentrations, reached a maximum of 0.65 µGy/h, which is relatively low considering background levels. No effects were observed on survival, fish condition, and metabolic indices (gonado-, hepato-, and spleno-somatic indexes (GSI, HSI, SSI), RNA/DNA and proteins/DNA ratios). Multivariate analyses showed that several biomarkers (DNA damages, micronucleus frequency, brain acetylcholinesterase, lysosomal membrane integrity, phagocytosis activity, and reactive oxygen species production) were exclusively correlated with fish tritium internal dose rate, showing that tritium induced genotoxicity, as well as neural and immune responses. The results were compared with another study on the same fish species where fish were exposed to tritium and other contaminants in natural environments. Together with the field study, the present work provides useful data to identify biomarkers for tritium exposure and better understand modes of action of tritium on the fathead minnow.

Cellular responses in rainbow trout (Oncorhynchus mykiss) reared in tritiated water and/or fed organically bound tritium.

For the evaluation of biological effects of low level chronic tritium exposure in fish, rainbow trout (Oncorhynchus mykiss) were exposed to tritiated water and/or fed a diet spiked with tritiated essential amino acids. Differences in fatty acid composition were noted following the tritium exposures. In addition, changes in response to a subsequent acute high dose of gamma radiation delivered in vitro were noted when evaluating DNA repair activity and fatty acid composition.

Effects of 90Sr on Tree Swallow Nestlings Near Groundwater Contaminant Plumes.

Discharge of groundwater contaminant plumes has created elevated concentrations of Sr in some aquatic sediments at Chalk River Laboratories. Tree swallows (Tachycenita bicolor) feed and supply their nestlings almost exclusively with airborne insects that developed as larvae in aquatic sediments. To monitor the uptake and test for potential detriment due to Sr in a terrestrial animal, we measured the gross beta concentrations in the bone of 12-d-old tree swallow nestlings in areas having sediments with elevated levels of gross beta (Sr and Y) and in several control areas where sediment gross beta was primarily due to naturally occurring K. Nesting behavior and reproductive success of the tree swallows were similar regardless of the gross beta concentrations in sediments near their nest boxes. Radiation can damage DNA and cause micronuclei to form in cells, so we examined the frequency of micronuclei in erythrocytes of nestlings. The formation of micronuclei in the erythrocytes of the nestlings was also similar wherever nestlings were analyzed. The results revealed no significant increases even near sediments with the highest gross beta levels. At Perch Lake, where Chalk River Laboratories has a large area of Sr-contaminated sediments, the bones of 12-d-old nestlings contained gross beta concentrations as high as 29 Bq g. This would produce a skeletal dose rate of 9 µGy h, which is one-fourth of the threshold dose rate of 40 µGy h, above which detriment could occur. Failing to find any indication of detriment in the field study, we irradiated wild eggs in the lab and returned them to their nest for natural incubation, hatching, and feeding by the parents. There was an increase in formation of micronuclei following a dose of 3.2 Gy, and the other results were consistent with existing literature.

Correlated responses for DNA damage, phagocytosis activity and lysosomal function revealed in a comparison between field and laboratory studies: Fathead minnow exposed to tritium.

Tritium entering the aquatic environment can confer a whole body internal radiological dose to aquatic organisms. Multiple stressors inherent in natural environments, however, confound estimates for observable radiation specific responses. To disentangle differences between field and laboratory outcomes to tritium exposures, a multivariate analysis comparing biomarkers for radiation exposure at the cellular level with changes in biological processes within tissues is described for fathead minnows (Pimephales promelas). Over tritium activity concentrations up to 180,000 Bq/L, DNA damage in the field were lower than DNA damage in the laboratory. This finding does not support an increase in morbidity of biota in field exposures. Energy deposited by tritium decay produces oxidised free radicals, yet the biological responses in brain, muscle and liver to oxidative stress differed between the studies and were not related to the tritium. For both studies, DNA damage in gonad and blood increased with increased tritium as did the fluorescence associated with lysosomal function in spleen. The studies differed in spleen phagocytosis activity were, in the laboratory but not the field, activity increased with increased tritium-and was correlatd with lysosomal function (Spearman coefficient of 0.98 (p = 0.001). The higher phagocytosis activity in the field reflects exposures to unmeasured factors that were not present within the laboratory. In the laboratory, DNA damage and lysosomal function were correlated: Spearman coefficients of 0.9 (Comet, p = 0.03) and 0.9 (micronuclei, p = 0.08). In the field, DNA damage by the Comet assay, but not by micronucleus frequency, correlated with lysosomal function: Spearman coefficients of 0.91 (Comet, p < 0.001) and 0.47 (micronuclei, p = 0.21). These observations highlight a need for better physiologic understanding of linkages between radiation-induced damage within cells and responses at higher levels of biological organization.

Spatial Autocorrelation, Source Water and the Distribution of Total and Viable Microbial Abundances within a Crystalline Formation to a Depth of 800 m.

Proposed radioactive waste repositories require long residence times within deep geological settings for which we have little knowledge of local or regional subsurface dynamics that could affect the transport of hazardous species over the period of radioactive decay. Given the role of microbial processes on element speciation and transport, knowledge and understanding of local microbial ecology within geological formations being considered as host formations can aid predictions for long term safety. In this relatively unexplored environment, sampling opportunities are few and opportunistic. We combined the data collected for geochemistry and microbial abundances from multiple sampling opportunities from within a proposed host formation and performed multivariate mixing and mass balance (M3) modeling, spatial analysis and generalized linear modeling to address whether recharge can explain how subsurface communities assemble within fracture water obtained from multiple saturated fractures accessed by boreholes drilled into the crystalline formation underlying the Chalk River Laboratories site (Deep River, ON, Canada). We found that three possible source waters, each of meteoric origin, explained 97% of the samples, these are: modern recharge, recharge from the period of the Laurentide ice sheet retreat (ca. ∼12000 years before present) and a putative saline source assigned as Champlain Sea (also ca. 12000 years before present). The distributed microbial abundances and geochemistry provide a conceptual model of two distinct regions within the subsurface associated with bicarbonate - used as a proxy for modern recharge - and manganese; these regions occur at depths relevant to a proposed repository within the formation. At the scale of sampling, the associated spatial autocorrelation means that abundances linked with geochemistry were not unambiguously discerned, although fine scale Moran's eigenvector map (MEM) coefficients were correlated with the abundance data and suggest the action of localized processes possibly associated with the manganese and sulfate content of the fracture water.

Biological effects of tritium on fish cells in the concentration range of international drinking water standards.

PURPOSE: To evaluate whether the current Canadian tritium drinking water limit is protective of aquatic biota, an in vitro study was designed to assess the biological effects of low concentrations of tritium, similar to what would typically be found near a Canadian nuclear power station, and higher concentrations spanning the range of international tritium drinking water standards. MATERIALS AND METHODS: Channel catfish peripheral blood B-lymphoblast and fathead minnow testis cells were exposed to 10-100,000 Bq l(-1) of tritium, after which eight molecular and cellular endpoints were assessed. RESULTS: Increased numbers of DNA strand breaks were observed and ATP levels were increased. There were no increases in γH2AX-mediated DNA repair. No differences in cell growth were noted. Exposure to the lowest concentrations of tritium were associated with a modest increase in the viability of fathead minnow testicular cells. Using the micronucleus assay, an adaptive response was observed in catfish B-lymphoblasts. CONCLUSIONS: Using molecular endpoints, biological responses to tritium in the range of Canadian and international drinking water standards were observed. At the cellular level, no detrimental effects were noted on growth or cycling, and protective effects were observed as an increase in cell viability and an induced resistance to a large challenge dose.

Health, growth and reproductive success of mice exposed to environmentally relevant levels of Ra-226 via drinking water over multiple generations.

PURPOSE: To assess health, growth and reproductive success of mammals exposed for multiple generations to levels of radium-226 known to occur in environments surrounding uranium mines and mills in Canada. METHODS: The study consisted of a control group and four treatment groups each containing 40 mice (20 males and 20 females) of the CBA/CaJ strain that were continuously exposed to a range of radium-226 levels via drinking water. Breeding was at 8-10 weeks of age and the study was concluded after three breeding cycles. RESULTS: When compared to control mice, constant consumption of drinking water containing 0.012, 0.076, 0.78 and 8.0 Bq/l of radium-226 over four generations of mice did not demonstrably affect physical condition, weight, pregnancy rate, number of pups per litter, sex ratio and bodyweight gain of pups. Between generations, the observed differences in pregnancy rates that were noted in all groups, including controls, seemed to directly correlate with the weight and age of the females at breeding. CONCLUSIONS: Based on the endpoints measured on four generations of mice, there is no indication that the consumption of radium-226 via drinking water (at activity concentrations up to 8.0 Bq/l) affects health, growth and reproductive fitness.