Cloning of Hynobius lichenatus (Tohoku hynobiid salamander) p53 and analysis of its expression in response to radiation.

BACKGROUND: Caudata species such as salamanders are easily affected by environmental changes, which can drastically reduce their population. The effects of acute X-rays and chronic γ-irradiation on Hynobius lichenatus, the Japanese Tohoku hynobiid salamander, are known. However, the expression of radiation-inducible genes, such as the DNA-damage checkpoint response gene p53, has not been analyzed in H. lichenatus. This has not occurred because there is no established method for mRNA quantification in H. lichenatus due to a lack of information on available nucleotide sequences corresponding to both radiation-inducible genes and endogenous control genes such as ACTB (ß-actin). RESULTS: In this study, we aimed to evaluate the effects of radiation on gene expression in H. lichenatus. Using RNA extracted from irradiated salamanders, we performed rapid amplification of cDNA ends (RACE) and cloned H. lichenatus ß-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and p53. We confirmed that the cloned cDNAs were able to synthesize salamander proteins by western blotting after transfection into cultured HEK293 cells. Proliferation assays using HEK293 cells stably expressing H. lichenatus p53 protein showed that this protein has antiproliferative effects, similar to that of mammalian p53. Furthermore, RT-qPCR analysis using gene-specific primers revealed that p53 mRNA expression in H. lichenatus was upregulated upon exposure to radiation. CONCLUSION: Our results suggest that H. lichenatus p53 protein take an important role in regulating the cellular responses to various stimuli as mammalian p53 does. Furthermore, our study provides novel data to select appropriate primers to analyze internal control mRNA expression in H. lichenatus and to evaluate p53 expression as a marker of radiation and environmental stimuli.

Chromosomal Aberrations in Large Japanese Field Mice (Apodemus speciosus) Captured near Fukushima Dai-ichi Nuclear Power Plant.

Since the Fukushima Dai-ichi Nuclear Power Plant accident, radiation effects on nonhuman biota in the contaminated areas have been a major concern. Here, we analyzed the frequencies of chromosomal aberrations (translocations and dicentrics) in the splenic lymphocytes of large Japanese field mice (Apodemus speciosus) inhabiting Fukushima Prefecture. A. speciosus chromosomes 1, 2, and 5 were flow-sorted in order to develop A. speciosus chromosome-specific painting probes, and FISH (fluorescence in situ hybridization) was performed using these painting probes to detect the translocations and dicentrics. The average frequency of the translocations and dicentrics per cell in the heavily contaminated area was significantly higher than the frequencies in the case of the noncontaminated control area and the slightly and moderately contaminated areas, and this aberration frequency in individual mice tended to roughly increase with the estimated dose rates and accumulated doses. In all four sampling areas, the proportion of aberrations occurring in chromosome 2 was approximately >3 times higher than that in chromosomes 1 and 5, which suggests that A. speciosus chromosome 2 harbors a fragile site that is highly sensitive to chromosome breaks induced by cellular stress such as DNA replication. The elevated frequency of chromosomal aberrations in A. speciosus potentially resulting from the presence of a fragile site in chromosome 2 might make it challenging to observe the mild effect of chronic low-dose-rate irradiation on the induction of chromosomal aberrations in A. speciosus inhabiting the contaminated areas of Fukushima.

Chromosomal Aberrations in Wild Mice Captured in Areas Differentially Contaminated by the Fukushima Dai-Ichi Nuclear Power Plant Accident.

Following the Fukushima Dai-ichi Nuclear Power Plant accident, radiation effects on nonhuman biota in the contaminated areas have been a great concern. The induction of chromosomal aberrations in splenic lymphocytes of small Japanese field mice (Apodemus argenteus) and house mice (Mus musculus) inhabiting Fukushima Prefecture was investigated. In mice inhabiting the slightly contaminated area, the average frequency of dicentric chromosomes was similar to that seen in mice inhabiting a noncontaminated control area. In contrast, mice inhabiting the moderately and heavily contaminated areas showed a significant increase in the average frequencies of dicentric chromosomes. Total absorbed dose rate was estimated to be approximately 1 mGy d(-1) and 3 mGy d(-1) in the moderately and heavily contaminated areas, respectively. Chromosomal aberrations tended to roughly increase with dose rate. Although theoretically, the frequency of chromosomal aberrations was considered proportional to the absorbed dose, chromosomal aberrations in old mice (estimated median age 300 days) did not increase with radiation dose at the same rate as that observed in young mice (estimated median age 105 days).

(135)Cs/(137)Cs isotopic ratio as a new tracer of radiocesium released from the Fukushima nuclear accident.

Since the Fukushima Daiichi nuclear power plant (FDNPP) accident in 2011, intensive studies of the distribution of released fission products, in particular (134)Cs and (137)Cs, in the environment have been conducted. However, the release sources, that is, the damaged reactors or the spent fuel pools, have not been identified, which resulted in great variation in the estimated amounts of (137)Cs released. Here, we investigated heavily contaminated environmental samples (litter, lichen, and soil) collected from Fukushima forests for the long-lived (135)Cs (half-life of 2 × 10(6) years), which is usually difficult to measure using decay-counting techniques. Using a newly developed triple-quadrupole inductively coupled plasma tandem mass spectrometry method, we analyzed the (135)Cs/(137)Cs isotopic ratio of the FDNPP-released radiocesium in environmental samples. We demonstrated that radiocesium was mainly released from the Unit 2 reactor. Considering the fact that the widely used tracer for the released Fukushima accident-sourced radiocesium in the environment, the (134)Cs/(137)Cs activity ratio, will become unavailable in the near future because of the short half-life of (134)Cs (2.06 years), the (135)Cs/(137)Cs isotopic ratio can be considered as a new tracer for source identification and long-term estimation of the mobility of released radiocesium in the environment.

Chromosomal Aberrations in Large Japanese Field Mice (Apodemus speciosus) Captured in Various Periods after Fukushima Daiichi Nuclear Power Plant Accident.

After the Fukushima Daiichi Nuclear Power Plant accident, we studied the chromosomal aberrations (dicentrics and translocations) in the splenic lymphocytes of wild mice inhabiting Fukushima prefecture. Here, we report the frequencies of chromosomal aberrations in large Japanese field mice (Apodemus speciosus) captured from 2012 to 2016 in a heavily contaminated area. The chromosomal aberrations were detected using newly developed 4-color FISH (fluorescence in situ hybridization) with A. speciosus chromosome 1-, 3-, 4- and 5-specific painting probes. The frequencies of chromosomal aberrations in mice captured in July 2012 and October 2014 were significantly higher than that in the mice inhabiting the non-contaminated control area; however, the frequency of chromosomal aberrations in mice captured in January 2016 was not. The frequency of chromosomal aberrations in individual mice tended to increase with certain dose rates and accumulated doses. Regression tree analyses suggested increasing chromosomal aberration rate in mice exposed to chronic radiation at dose rates of more than 1.1 mGy day-1 and at accumulated doses of more than 200 mGy. It is concluded that ambient dose rates in the most severely contaminated area of Fukushima prefecture and radiation doses to wild mice inhabiting this area decrease with time; consequently, chromosomal aberrations induced by radiation have not been detected 5 years after the accident.

Genome-wide gene expression profiling in gamma-irradiated green alga, Pseudokirchneriella subcapitata, by HiCEP.

Transcriptome was analyzed in gamma-irradiated green alga Pseudokirchneriella subcapitata by high coverage gene expression profiling (HiCEP). Approximately 7,800 expressed genes were detected. Expression levels of 623-707 genes were affected at 100-300 Gy. Nucleotide sequences of 41 up-regulated genes were determined. The quantitative reverse transcription polymerase chain reaction validated the up-regulation. Two genes had homology to genes related to ionizing radiation. These results indicate usefulness of HiCEP for screening of stress-responsive genes in species that are ecotoxicologically important but for which genomic sequence information is lacking.

Dose rate estimation of freshwater wildlife inhabiting irrigation ponds in the exclusion zone of the Fukushima Dai-ichi Nuclear Power Plant accident.

To assess the risks of ionising radiation to freshwater environments in the exclusion zone of the Fukushima Dai-ichi Nuclear Power Plant accident, the absorbed dose rates to aquatic organisms possibly inhabiting the irrigation ponds were estimated using the ERICA Assessment Tool from 134Cs and 137Cs radioactivity monitoring data for the period 2013 to 2017. In each year, the total dose rates to benthic organisms were in the same or higher levels compared with those to pelagic organisms. Among pelagic organisms, the total dose rates to amphibians, birds, and pelagic fish were two orders of magnitude higher than those to plankton. The total dose rates to insect larvae, which attained a maximum of 130 µGy h-1, were higher than those to the other benthic organisms. The dose rates to benthic organisms increased from 2013 to 2015 and remained constant thereafter. In 50-93% of ponds, the dose rates to at least one taxon of freshwater organism, all of which were benthic organisms, exceeded the ERICA screening level (10 µGy h-1). Comparison of the estimated dose rates with the ICRP's derived consideration reference levels (DCRLs) suggests that radioactive contamination was not likely to damage amphibians, birds, pelagic fish, benthic fish, crustaceans, and insect larvae inhabiting most of the irrigation ponds in the exclusion zone. However, this comparison also suggests that there was some chance of deleterious effects occurring to birds and benthic fish in a limited number of the most severely contaminated irrigation ponds.

Effects of chronic γ-irradiation on growth and sexual maturation of the Tohoku hynobiid salamander, Hynobius lichenatus.

There are still considerable gaps in knowledge regarding the biological effects of chronic ionising radiation exposure in amphibians. To fill these gaps, Tohoku hynobiid salamanders, Hynobius lichenatus (Amphibia, Caudata), were chronically irradiated with 137Cs γ-rays from embryonic to adult stages over 1954 days, and the effects on their growth and sexual maturation were examined under laboratory conditions. Irradiation at a dose rate of 33 µGy h-1 had some stimulatory effects on growth (body weight increase) of H. lichenatus, while growth was temporarily or permanently suppressed at 150 or 510 µGy h-1, respectively. On day 1802, secondary sexual characteristics (a tubercle at the anterior angle of the cloacal vent for males and ovisac development for females) were observed in 91% of the salamanders irradiated at 33 µGy h-1, and in a similar percentage of non-irradiated controls. At 150 and 510 µGy h-1, secondary sexual characteristics were not observed in any individuals. These results suggest that the derived consideration reference level (DCRL) of the International Commission on Radiological Protection (ICRP) for Reference Frog, i.e. 40-400 µGy h-1, is applicable for the protection of H. lichenatus, and that growth and sexual maturation of this salamander may not have been adversely affected even in the most severely contaminated area in Fukushima, where the highest dose rate to salamanders was estimated to be 50 µGy h-1. However, observations in the contaminated area are required to confirm this conclusion, considering the possible confounding factors which may make this salamander more sensitive to radiation in the natural environment than under laboratory conditions.

Impact of gamma irradiation on the transformation efficiency for extracellular plasmid DNA.

Extracellular DNA is omnipresent in aquatic environments and is thought to be a genetic material for horizontal gene transformation between microorganisms. We studied the impact of gamma irradiation on the transformation efficiency (transformants number per ng of DNA per ml) of extracellular DNA. Plasmid pEGFP as a model extracellular DNA was irradiated by gamma rays. The transformation efficiency decreased with the increase in radiation dose. A total dose of 10Gy is normally not lethal for microorganisms but certainly affects the transformation efficiency of extracellular DNA. The decrease in the efficiency would be induced by strand breaks of extracellular DNA because the yield of both single-strand breaks (SSBs) and double-strand breaks (DSBs) increased with the increase in radiation dose. The relative transformation efficiency of SSBs and DSBs to that of covalently closed circles (CCCs) was 30.3% and 0.2%, respectively. This impact on natural transformation suggests an inability of microorganisms to acquire new characteristics which should be normally acquired.

Radiocaesium contamination and dose rate estimation of terrestrial and freshwater wildlife in the exclusion zone of the Fukushima Dai-ichi Nuclear Power Plant accident.

To characterise the radioactive contamination of terrestrial and freshwater wildlife caused by the Fukushima Dai-ichi Nuclear Power Plant accident, biological samples, namely, fungi, mosses, plants, amphibians, reptiles, insects, molluscs, and earthworms, were collected mainly from the forests of the exclusion zone in the Fukushima Prefecture from 2011 to 2012. Caesium-134 and 137Cs were detected by gamma spectrometry in almost all the samples. Fungi, ferns, and mosses accumulated high amounts of radiocaesium, as they did in Chernobyl, with 134Cs + 137Cs activity concentrations of 104-106 Bq kg-1 fresh mass (FM). Earthworms, amphibians, and the soft tissue of the garden snail Acusta despecta sieboldiana, also had levels as high as 104-105 Bq kg-1 FM of 134Cs + 137Cs. Most of the estimated total (internal + external) dose rates to herbaceous plants, amphibians, insects, and earthworms were below the corresponding derived consideration reference levels (DCRLs) recommended by the ICRP. This suggests that, in most cases, there was little chance of deleterious effects of ionising radiation on these organisms in the exclusion zone for the first year after the accident, though the dose rates were underestimated mainly due to the lack of consideration of short-lived radionuclides.

Radiocaesium contamination of wild boars in Fukushima and surrounding regions after the Fukushima nuclear accident.

Analysis of radioactivity data obtained under the food monitoring campaign in Japan indicates that elevated 134Cs+137Cs activity concentrations in wild boar meat remained constant or slowly decreased in Fukushima and surrounding prefectures from 2011 to 2015. The activity concentrations in some samples are still over the regulatory limit of 100 Bq kg-1 fresh weight, even in 2015. Activity concentrations of 137Cs in muscle of wild boars we captured in 2011 were higher than those in kidney, liver, spleen, heart and lung. A food processing retention factor, Fr, was 0.5 or 0.6 for 137Cs when the wild boar meat was boiled, suggesting that a parboiling process is effective for reduction of radiocaesium intake from wild boar meat.

Morphological defects in native Japanese fir trees around the Fukushima Daiichi Nuclear Power Plant.

After the accident at the Fukushima Daiichi Nuclear Power Plant (F1NPP) in March 2011, much attention has been paid to the biological consequences of the released radionuclides into the surrounding area. We investigated the morphological changes in Japanese fir, a Japanese endemic native conifer, at locations near the F1NPP. Japanese fir populations near the F1NPP showed a significantly increased number of morphological defects, involving deletions of leader shoots of the main axis, compared to a control population far from the F1NPP. The frequency of the defects corresponded to the radioactive contamination levels of the observation sites. A significant increase in deletions of the leader shoots became apparent in those that elongated after the spring of 2012, a year after the accident. These results suggest possibility that the contamination by radionuclides contributed to the morphological defects in Japanese fir trees in the area near the F1NPP.

Estimation of absorbed radiation dose rates in wild rodents inhabiting a site severely contaminated by the Fukushima Dai-ichi nuclear power plant accident.

The dose rates of radiation absorbed by wild rodents inhabiting a site severely contaminated by the Fukushima Dai-ichi Nuclear Power Plant accident were estimated. The large Japanese field mouse (Apodemus speciosus), also called the wood mouse, was the major rodent species captured in the sampling area, although other species of rodents, such as small field mice (Apodemus argenteus) and Japanese grass voles (Microtus montebelli), were also collected. The external exposure of rodents calculated from the activity concentrations of radiocesium ((134)Cs and (137)Cs) in litter and soil samples using the ERICA (Environmental Risk from Ionizing Contaminants: Assessment and Management) tool under the assumption that radionuclides existed as the infinite plane isotropic source was almost the same as those measured directly with glass dosimeters embedded in rodent abdomens. Our findings suggest that the ERICA tool is useful for estimating external dose rates to small animals inhabiting forest floors; however, the estimated dose rates showed large standard deviations. This could be an indication of the inhomogeneous distribution of radionuclides in the sampled litter and soil. There was a 50-fold difference between minimum and maximum whole-body activity concentrations measured in rodents at the time of capture. The radionuclides retained in rodents after capture decreased exponentially over time. Regression equations indicated that the biological half-life of radiocesium after capture was 3.31 d. At the time of capture, the lowest activity concentration was measured in the lung and was approximately half of the highest concentration measured in the mixture of muscle and bone. The average internal absorbed dose rate was markedly smaller than the average external dose rate (<10% of the total absorbed dose rate). The average total absorbed dose rate to wild rodents inhabiting the sampling area was estimated to be approximately 52 µGy h(-1) (1.2 mGy d(-1)), even 3 years after the accident. This dose rate exceeds 0.1-1 mGy d(-1) derived consideration reference level for Reference rat proposed by the International Commission on Radiological Protection (ICRP).

Dose rate estimation of the Tohoku hynobiid salamander, Hynobius lichenatus, in Fukushima.

The radiological risks to the Tohoku hynobiid salamanders (class Amphibia), Hynobius lichenatus due to the Fukushima Dai-ichi Nuclear Power Plant accident were assessed in Fukushima Prefecture, including evacuation areas. Aquatic egg clutches (n = 1 for each sampling date and site; n = 4 in total), overwintering larvae (n = 1-5 for each sampling date and site; n = 17 in total), and terrestrial juveniles or adults (n = 1 or 3 for each sampling date and site; n = 12 in total) of H. lichenatus were collected from the end of April 2011 to April 2013. Environmental media such as litter (n = 1-5 for each sampling date and site; n = 30 in total), soil (n = 1-8 for each sampling date and site; n = 31 in total), water (n = 1 for each sampling date and site; n = 17 in total), and sediment (n = 1 for each sampling date and site; n = 17 in total) were also collected. Activity concentrations of (134)Cs + (137)Cs were 1.9-2800, 0.13-320, and 0.51-220 kBq (dry kg) (-1) in the litter, soil, and sediment samples, respectively, and were 0.31-220 and <0.29-40 kBq (wet kg)(-1) in the adult and larval salamanders, respectively. External and internal absorbed dose rates to H. lichenatus were calculated from these activity concentration data, using the ERICA Assessment Tool methodology. External dose rates were also measured in situ with glass dosimeters. There was agreement within a factor of 2 between the calculated and measured external dose rates. In the most severely contaminated habitat of this salamander, a northern part of Abukuma Mountains, the highest total dose rates were estimated to be 50 and 15 µGy h(-1) for the adults and overwintering larvae, respectively. Growth and survival of H. lichenatus was not affected at a dose rate of up to 490 µGy h(-1) in the previous laboratory chronic gamma-irradiation experiment, and thus growth and survival of this salamander would not be affected, even in the most severely contaminated habitat in Fukushima Prefecture. However, further studies of the adult salamanders may be required in order to examine whether the most severe radioactive contamination has any effects on sensitive endpoints, since the estimated highest dose rate to the adults exceeded some of the guidance dose rates proposed by various organisations and programmes for the protection of amphibians, which range from 4 to 400 µGy h(-1). Conversely, at one site in Nakadori, a moderately contaminated region in Fukushima Prefecture, the dose rate to the adult salamanders in spring of 2012 was estimated to be 0.2 µGy h(-1). Estimated dose rates to the overwintering larvae in spring of 2012 were 1 and 0.2 µGy h(-1) at one site in Nakadori, and in Aizu, a less contaminated region in Fukushima Prefecture, respectively. These results suggest that there is a low risk that H. lichenatus will be affected by radioactive contamination in these districts, though further studies on dose rate estimation are required for definitive risk characterisation.

Release of transforming plasmid DNA from actively growing genetically engineered Escherichia coli.

We studied the transforming ability of the extracellular plasmid DNA released from a genetically engineered Escherichia coli pEGFP and the culturing conditions for the release of transforming DNA. The transforming ability was evaluated by transformation of competent cells with filtrates of E. coli pEGFP cultures. The number of transformants increased with time when E. coli pEGFP cells grew exponentially in rich medium, but not in stationary phase or when inoculated in freshwater. These results suggested that crude extracellular plasmid DNA had transforming ability and this transforming DNA was mainly released by actively growing bacteria.

Protection of negative gravitaxis in Euglena gracilis Z against gamma-ray irradiation by Trolox C.

The protective effects of Trolox on the inhibition of negative gravitaxis in Euglena gracilis exposed to 200 Gy 60Co gamma-rays were examined using different concentrations (1, 10 and 100 microM). The orientation precision of the negative gravitaxis was quantified using the r-value. A significant decrease in the r-value was observed in gamma-irradiated samples (0.18+/-0.03) compared to those of non-irradiated samples (0.47+/-0.03). There were no significant changes in the r-value of cells exposed to 200 Gy gamma-rays by the addition of 1 or 10 microM of Trolox. A significant increase (0.19) in the r-value of cells exposed to 200 Gy with 100 microM Trolox was observed. The results indicates that Trolox at a concentration of 100 microM protects negative gravitaxis against 60Co gamma-ray irradiation at a dose of 200 Gy. It also suggests that the negative gravitaxis of Euglena gracilis is affected by free radicals.

Cytokinesis block micronucleus assay in field plants for monitoring radiation-induced genotoxicity of the environment.

Effective biomonitoring for detection of radiation-induced genotoxicity of contaminants in natural environments involves testing of field plants for cytogenetic changes. To increase the efficiency and precision of cytogenetic analyses of field plants that have naturally high individual variability, an improved micronucleus assay is proposed that employs a cytokinesis block technique similar to the lymphocyte test system used in mammals. In seed embryonic meristems of the Japanese cedar, application of a methylxanthine derivative, 3-isobutyl-1-methylxanthine (IBMX), was found to be effective in inhibiting cytokinesis to make once-divided cells easily recognizable by their binucleate appearance. In the meristem of IBMX-treated seminal roots from X-ray-irradiated seeds, variation in micronucleus frequency in the binucleate cell population was reduced compared to that in the total cell population. The highest efficiency of measurement of micronucleus frequencies was obtained in the root meristems where 0.2- to 1.5-mm-long seminal roots were incubated with IBMX for 24 h. This result indicated that this root elongation stage corresponded to the first divisions of the root meristematic cells, and was therefore suitable for obtaining reliable estimations of accumulated genetic damage in the seeds. This cytokinesis block assay applied specifically at the root elongation stage was then used to examine dose-response relationships in Japanese cedar seeds irradiated either acutely with X-rays or chronically with γ-rays. The resulting dose-response curve for the acute X-ray irradiation was fitted onto a linear-quadratic regression curve, whereas the dose-response curve for the chronic γ-irradiation matched a linear regression line better. Both dose-response curves were consistent with the target theory of classical radiation biology. The good agreement of the micronucleus data to a simple dose-response model indicates the proposed accuracy of the cytokinesis block micronucleus assay for plant monitoring.

Effects of chronic γ-irradiation on growth and survival of the Tohoku hynobiid salamander, Hynobius lichenatus.

The Tohoku hynobiid salamanders, Hynobius lichenatus, were chronically irradiated with γ-rays from embryonic to juvenile stages for 450 days. At 490 µGy h(-1) or lower dose rates, growth and survival were not significantly affected by irradiation, and any morphological aberrations and histological damages were not observed. At 4600 µGy h(-1), growth was severely inhibited, and all the individuals died mostly at the juvenile stage. Chronic LD50 was 42 Gy as a total dose. In the liver, the number of hematopoietic cells was significantly reduced in the living juveniles, and these cells disappeared in the dead juveniles. In the spleen, mature lymphocytes were depleted in the living larvae, and almost all the heamtopoietic cells disappeared in the dead juveniles. These results suggest that this salamander died due to acute radiation syndrome, i.e., hematopoietic damage and subsequent sepsis caused by immune depression. The death would be also attributed to skin damage inducing infection. At 18,000 µGy h(-1), morphological aberrations and severe growth inhibition were observed. All the individuals died at the larval stage due to a multiple organ failure. Chronic LD50 was 28 Gy as a total dose. Assuming that chronic LD50 was 42 Gy at lower dose rates than 4600 µGy h(-1), a chronic median lethal dose rate could be estimated to be <340 µGy h(-1) for the whole life (>14 years). These results suggest that, among guidance dose rates, i.e., 4-400 µGy h(-1), proposed by various organisations and research programmes for protection of amphibians and taxonomic groups or ecosystems including amphibians, most of them would protect this salamander but the highest value may not on the whole life scale.

Effects of chronic γ-irradiation on the aquatic microbial microcosm: equi-dosimetric comparison with effects of heavy metals.

Effects of chronic γ-irradiation were investigated in the aquatic microcosm consisting of flagellate algae Euglena gracilis as producers, ciliate protozoa Tetrahymena thermophila as consumers and bacteria Escherichia coli as decomposers. At 1.1 Gy day(-1), no effects were observed. At 5.1 Gy day(-1), cell densities of E. coli showed a tendency to be lower than those of controls. At 9.7 and 24.7 Gy day(-1), population decrease was observed in E. coli. E. gracilis and T. thermophila died out after temporal population decrease and subsequent population increase in T. thermophila. It is likely that this temporal population increase was an indirect effect due to interspecies interactions. Effect dose rates of γ-rays were compared with effect concentrations of some metals using the radiochemoecological conceptual model and the effect index for microcosm. Comparison of these community-level effects data with environmental exposure data suggests that ionising radiation, gadolinium and dysprosium have low risks to affect aquatic microbial communities while manganese, nickel and copper have considerable risks. Effects of chronic irradiation were smaller than those of acute irradiation, and an acute to chronic ratio was calculated to be 28 by dividing an acute dose by chronic daily dose rate at which the effect index was 10%. This ratio would be useful for community-level extrapolation from acute to chronic radiation effects.

Isotopic evidence of plutonium release into the environment from the Fukushima DNPP accident.

The Fukushima Daiichi nuclear power plant (DNPP) accident caused massive releases of radioactivity into the environment. The released highly volatile fission products, such as (129m)Te, (131)I, (134)Cs, (136)Cs and (137)Cs were found to be widely distributed in Fukushima and its adjacent prefectures in eastern Japan. However, the release of non-volatile actinides, in particular, Pu isotopes remains uncertain almost one year after the accident. Here we report the isotopic evidence for the release of Pu into the atmosphere and deposition on the ground in northwest and south of the Fukushima DNPP in the 20-30 km zones. The high activity ratio of (241)Pu/(239+240)Pu (> 100) from the Fukushima DNPP accident highlights the need for long-term (241)Pu dose assessment, and the ingrowth of (241)Am. The results are important for the estimation of reactor damage and have significant implication in the strategy of decontamination.