New data on mobility of transuranium elements in sediments of the Yenisei River.

The inflow of transuranium elements to the Yenisei River was previously associated with the production of weapons-grade plutonium at the Mining-and-Chemical Combine (MCC, Zheleznogorsk, Russia), but the source of transuranium elements in the River today is fabrication of MOX fuel that started recently at the MCC. The current study presents results of sequential chemical extraction of radionuclides from sediment samples collected in 2014 and 2020 in two areas near the MCC discharge site and compares these results with the data obtained previously by sequential extraction of sediments collected during 1999-2007. Over the study period, the strength of binding of 137Cs and 60Co in the Yenisei River sediments was high (up to 100%) and remained so, while the percentages of 241Am and 152Eu in residual solids after sequential extraction increased considerably and the percentage of 239,240Pu in residual solids decreased in samples from all study areas. In samples collected at the position located close to the MCC discharge site, the percentages of the strongly bound 241Am and 152Eu as well as 239,240Pu were lower than in the samples from the other positions. The study demonstrated an enormous increase in 239,240Pu activity concentration in the top sediment layers collected at all positions in 2020 relative to 2014. In the same period, as literature data suggest, 239,240Pu activity concentrations also increased in aquatic organisms of the Yenisei River, which can be indicative of the growing potential bioavailability of plutonium in the aquatic ecosystem, which could be caused by the presence of the mobile form of plutonium in the routine discharges from the MCC.

The use of the aquatic plant Elodea canadensis to assess the effects of low-dose gamma irradiation.

Responses of cytogenetic and growth endpoints of the aquatic plant Elodea canadensis were studied using the plants collected from non-contaminated control areas of the Yenisei River and exposed to external γ-radiation for 11-13 days in the laboratory. Elodea canadensis was exposed to 0.5-25 mGy/day γ-radiation dose rates from a 137Cs source. The total number of aberrant cells and total root length were more sensitive elodea endpoints to γ-radiation than total shoot length and mitotic index of elodea. Radiation sensitivity of elodea can be compared with the sensitivity of such a Reference Plant as Wild Grass (1-10 mGy/day) recommended by the ICRP. Thus, the aquatic plant Elodea canadensis can be used as a radiation bioindicator.

The long-term effects of γ-radiation on the growth of Allium cepa plants.

PURPOSE: The purpose of this study was to assess the long-term effects of gamma-radiation, including low-dose radiation, on growth parameters of onion (Allium cepa) seedling roots 6-10 days after irradiation. MATERIALS AND METHODS: Onion seedlings were exposed to a 137Cs gamma source at doses ranging from 0.1 to 10 grays (Gy). Responses of root and shoot length growth were studied 6 and 10 days after irradiation. RESULTS: Our results showed inhibition of the root and shoot length growth 6 days after exposure at all doses, including the low dose - 0.1 Gy. At a later point in time (day 10), root and shoot inhibition was only observed after irradiation at high doses (above 5 Gy), and that suggested the occurrence of cell repair after irradiation at low doses. The results indicated that the length of seedling roots was more sensitive to gamma-irradiation than the shoot length. CONCLUSION: The results of the study suggested that short-term gamma-irradiation of onion seedlings (absorbed doses of 0.1-10 Gy) caused inhibition of plant growth 6 and 10 days after irradiation. The dose dependence of the onion root length was linear. The present study showed for the first time that short-term low-dose gamma-irradiation could induce long-term negative effects on plant growth.

A long-term study of radionuclide concentrations in mushrooms in the 30-km zone around the Mining-and-Chemical Combine (Russia).

137Cs concentrations were measured in mushrooms in an area affected by radioactive discharges of the Mining-and-Chemical Combine (MCC) (Rosatom, Russia) in 2002-2017. The sources of radionuclides in the study sites were global fallouts and waterborne and airborne radioactive discharges of the MCC. The mushroom species Suillus granulatus and S. luteus showed the highest 137Cs concentrations (140-7100 Bq kg-1) for this area. Over the entire monitoring period, no significant change in 137Cs concentration was observed in the Suillus spp. samples collected from the sites with the aerial deposition of radionuclides. In the floodplain site with the radionuclide deposition from water and air, a significant decrease in the average 137Cs concentration was observed in the period between 2004 and 2017: a three-fold decrease in Suillus spp. and a nine-fold decrease in Lactarius deliciosus. The effective half-lives of 137Cs in fruiting bodies of the mushrooms L. deliciosus and Suillus spp. in this site were 3.6 ± 0.6 and 9.2 ± 2.7 years, respectively.

Biomonitoring of radioactive contamination of the Yenisei River using aquatic plants.

The Yenisei River is contaminated by artificial radionuclides released by one of the Russian facilities producing weapons-grade plutonium (the Mining-and-Chemical Combine, MCC), which has been in operation for 60 years. The paper presents results of long-term monitoring of radionuclide concentrations in six aquatic plant species collected from the 1400 km Yenisei River stretch downstream of the city of Krasnoyarsk. Before the last MCC reactor was shut down (in 2010), up to 30 artificial radionuclides were detected in the plant biomass, and 2 and 5 years after the reactor shutdown, 11 and 3-5 radionuclides, respectively, were detected. The highest concentrations of radionuclides were recorded in Fontinalis antipyretica. The aquatic plant Elodea canadensis, which commonly occurs in the Yenisei River, was used in the cytogenetic study. High frequencies of chromosomal aberrations (up to 33%) were revealed in cells of Elodea canadensis roots from the Yenisei region affected by the MCC radioactive discharge, at dose rates of 45-72 µGy/d, while in the cells of the plant roots from the reference areas, the frequency of chromosomal aberrations was 5-7%, at dose rates below 2 µGy/d. The higher frequencies of chromosomal aberrations in Elodea canadensis collected from the parts of the River with increased concentrations of artificial radionuclides are associated with the radiation factor. The aquatic plant Elodea canadensis can be recommended as a bio-indicator for radioactively contaminated aquatic ecosystems.

Radioactive particles in the Yenisei River floodplain (Russia): Characterization, leaching and potential effects in the environment.

The operation of the Mining-and-Chemical Combine (MCC), the largest producer of weapons-grade plutonium in Russia, has resulted in radioactive contamination of the Yenisei River floodplain. Investigations carried out in Novosibirsk and Krasnoyarsk institutes have shown that the floodplain of the Yenisei downstream of the MCC is contaminated by radioactive particles (RP) of various types and activities. Analytical characterization of the RP showed that most of them were fuel particles, which were carried into the Yenisei after incidents at the MCC reactors. The plutonium and caesium isotope ratios (238Pu/239,240Pu; 137Cs/134Cs) vary substantially between the particles, indicating different source terms and time intervals when the RP were formed. In addition to fuel RP, there were particles that contained activation radionuclides. The experiment on dissolution of RP using the model solution (the simulated stomach fluid) showed different cumulative extractions of radionuclides from the particles: 60Co and 137Cs extractions were the lowest, the extracted fractions of europium and americium isotopes were the largest, and plutonium occupied an intermediate position. High concentrations of radionuclides in RP are sources of exposure of organisms in aquatic and terrestrial ecosystems to low radiation doses. The plant bioassays of the effects of γ-radiation from RP showed the effect of low doses of γ-radiation on growth parameters of aquatic plant Elodea canadensis growing in the Yenisei River floodplain. The presence of RP from different sources in the Yenisei River floodplain makes this region a unique site for studying environmental effects of the particles.

Protein-based fluorescent bioassay for low-dose gamma radiation exposures.

The study suggests an application of a coelenteramide-containing fluorescent protein (CLM-CFP) as a simplest bioassay for gamma radiation exposures. "Discharged obelin," a product of the bioluminescence reaction of the marine coelenterate Obelia longissima, was used as a representative of the CLM-CFP group. The bioassay is based on a simple enzymatic reaction-photochemical proton transfer in the coelenteramide-apoprotein complex. Components of this reaction differ in fluorescence color, providing, by this, an evaluation of the proton transfer efficiency in the photochemical process. This efficiency depends on the microenvironment of the coelenteramide within the protein complex, and, hence, can evaluate a destructive ability of gamma radiation. The CLM-CFP samples were exposed to gamma radiation (137Cs, 2 mGy/h) for 7 and 16 days at 20 °C and 5 °C, respectively. As a result, two fluorescence characteristics (overall fluorescence intensity and contributions of color components to the fluorescence spectra) were identified as bioassay parameters. Both parameters demonstrated high sensitivity of the CLM-CFP-based bioassay to the low-dose gamma radiation exposure (up to 100 mGy). Higher temperature (20 °C) enhanced the response of CLM-CFP to gamma radiation. This new bioassay can provide fluorescent multicolor assessment of protein destruction in cells and physiological liquids under exposure to low doses of gamma radiation. Graphical abstract ᅟ.

Unique diversity of radioactive particles found in the Yenisei River floodplain.

The long-term operation of three reactors and the radiochemical plant of the Mining-and-Chemical Combine (MCC), Russia's largest producer of weapons-grade plutonium, has resulted in radioactive contamination of the Yenisei River floodplain. From 1995 to 2016, we found more than 200 radioactive particles (RP) in the Yenisei floodplain, downstream of the MCC. Analytical characterization showed that most of the RP were fuel particles, which were carried into the river after incidents at the MCC reactors. Having compared the 137Cs/134Cs ratios in the particles, we determined three time intervals when the RP were formed. The plutonium isotope ratios (238Pu/239,240Pu) vary substantially between the particles and indicate several different source terms. In addition to fuel RP, we found particles that only contained activation products (60Co or europium isotopes). SEM and γ-spectrometry showed that the cobalt particles could have originated from the corrosion of the reactor coolant system and the europium particles - from the damaged compensating rods. No europium particles have been found anywhere else in the world. The presence of RP from different sources (fuel, cobalt, and europium particles) in the Yenisei River floodplain makes this region a unique site for studying environmental effects of the particles. These RP represent point sources of radioecological significance.

Low doses of gamma-radiation induce SOS response and increase mutation frequency in Escherichia coli and Salmonella typhimurium cells.

This study addresses use of two bacterial test systems (the Ames test and the SOS chromotest) to estimate the effects of low doses of γ-radiation. The most substantial increases in induction of SOS response and mutation frequencies were observed in the first 24h of exposure to γ-radiation as compared to the cells in the exposure-free control. Gamma-radiation also impaired growth and survival of S. typhimurium cells in the first 24h. The effects were attenuated at lower exposure doses and at longer exposure times. In the experiments conducted in this study, at 96h of exposure, the values of some of the γ-radiation effects were lower than the MID (minimum inducing dose) detection limits and, thus, were neglected. Long-term exposure to γ-radiation could also result in combined effects of γ-radiation and the death of cells in the culture.

Chromosomal abnormalities in roots of aquatic plant Elodea canadensis as a tool for testing genotoxicity of bottom sediments.

Submersed freshwater macrophytes are considered as relevant indicators for use in bulk bottom sediment contact tests. The purpose of this study was to estimate the validity of endpoints of aquatic plant Elodea canadensis for laboratory genotoxicity testing of natural bottom sediments. The inherent level of chromosome abnormalities (on artificial sediments) in roots of E. canadensis under laboratory conditions was lower than the percentage of abnormal cells in bulk sediments from the Yenisei River. The percentage of abnormal cells in roots of E. canadensis was more sensitive to the presence of genotoxic agents in laboratory contact tests than in the natural population of the plant. The spectra of chromosomal abnormalities that occur in roots of E. canadensis under natural conditions in the Yenisei River and in laboratory contact tests on the bulk bottom sediments from the Yenisei River were similar. Hence, chromosome abnormalities in roots of E. canadensis can be used as a relevant and sensitive genotoxicity endpoint in bottom sediment-contact tests.

Use of the aquatic plant Elodea canadensis to assess toxicity and genotoxicity of Yenisei River sediments.

The toxicity, cytotoxicity, and genotoxicity of bulk sediments from the Yenisei River (Siberia, Russia) were estimated in laboratory bioassays based on several endpoints in the aquatic plant Elodea canadensis. The bottom sediment samples were collected in the Yenisei River upstream and downstream of the sources of chemical and radioactive contamination. The testing revealed different sensitivities of Elodea endpoints to the quality of the bottom sediment: weight of shoots < length of shoots < mitotic index < length of roots < percentage of abnormal cells. The response of the genotoxicity endpoint (percentage of cells with chromosome abnormalities in roots of Elodea) was the highest in sediments with chemical pollution, whereas the highest inhibition of toxicity endpoints (shoot and root length) occurred in sediments with the highest level of radioactive pollution. The extreme response of Elodea endpoints to the quality of certain sediment samples may be regarded as related to the possible presence of unknown toxicants. The results show that E. canadensis can be used as an indicator species in laboratory contact testing of bottom sediment. The responses of shoot and root length growth endpoints of Elodea can be recommended as basic sensitivity indicators of bottom sediment toxicity. Analysis of cells carrying abnormal chromosomes in the apical root meristem of Elodea can be performed optionally in the same test to assess the genotoxicity of sediments.

Chemical fractionation of radionuclides and stable elements in aquatic plants of the Yenisei River.

The Yenisei River is contaminated with artificial radionuclides released by one of the Russian nuclear plants. The aquatic plants growing in the radioactively contaminated parts of the river contain artificial radionuclides. The aim of the study was to investigate accumulation of artificial radionuclides and stable elements by submerged plants of the Yenisei River and estimate the strength of their binding to plant biomass by using a new sequential extraction scheme. The aquatic plants sampled were: Potamogeton lucens, Fontinalis antipyretica, and Batrachium kauffmanii. Gamma-spectrometric analysis of the samples of aquatic plants has revealed more than 20 radionuclides. We also investigated the chemical fractionation of radionuclides and stable elements in the biomass and rated radionuclides and stable elements based on their distribution in biomass. The greatest number of radionuclides strongly bound to biomass cell structures was found for Potamogeton lucens and the smallest for Batrachium kauffmanii. For Fontinalis antipyretica, the number of distribution patterns that were similar for both radioactive isotopes and their stable counterparts was greater than for the other studied species. The transuranic elements (239)Np and (241)Am were found in the intracellular fraction of the biomass, and this suggested their active accumulation by the plants.

Detoxification of AM-241 solutions by humic substances: bioluminescent monitoring.

The study addresses the effect of humic substances on marine luminous bacteria Photobacterium phosphoreum exposed to Am-241 (3,000 Bq L(-1), water solution). Luminescent intensity of the bacteria was applied as a marker of their physiological activity. Humic substances have been found to reduce the effect of Am-241 on luminescence, decrease damage to cells, and change distribution of Am-241 between bacterial cells and intercellular media. It was shown that water-soluble humic substances, being products of natural transformation of organic substances in soil and bottom sediments, can serve as protecting agents for water microorganisms exposed to alpha radionuclides.

Effect of low-level alpha-radiation on bioluminescent assay systems of various complexity.

This study addresses the effects of low-level alpha-radiation on bioluminescent assay systems of different levels of organization: in vivo and in vitro. Three bioluminescent assay systems are used: intact bacteria, lyophilized bacteria, and bioluminescent system of coupled enzyme reactions. Solutions of 241Am(NO3)3 are used as a source of alpha-radiation. It has been shown that activation processes predominate in all the three bioluminescent assay systems subjected to short-term exposure (20-55 h) and inhibition processes in the systems subjected to longer-term exposure to radiation. It has been found that these effects are caused by the radiation component of 241Am3+ impact. The intensity of the 241Am3+ effect on the bioluminescent assay systems has been shown to depend on the 241Am3+ concentration, level of organization and integrity of the bioluminescent assay system. The bioluminescent assay systems in vivo have been found to be highly sensitive to 241Am3+ (up to 10(-17) M).