Prussian Blue to reduce radiocaesium accumulation in fish in lakes affected by the Chornobyl accident.

Fish in lakes situated within the Chornobyl exclusion zone have been highly contaminated since the accident and have not been utilized as food for humans. In the present study, field experiments with less-contaminated silver Prussian carp (Carassius gibelio (Bloch, 1782)) caged in contaminated lake within the Chornobyl exclusion zone was performed from June to October 2021 to investigate the effectiveness of clean feed containing potassium ferric hexacyanoferrate (KFCF) KFe[Fe(CN)6], a kind of Prussian Blue as a countermeasure to reduce 137Cs accumulation in fish. The addition of clean feed containing 0.1% or 1% KFCF resulted respectively in 2.4 ± 0.4 or 4.2 ± 0.7 times lower activity concentration of 137Cs in muscle tissue of the carp compared to control fish with clean feed without KFCF and in 7-16 or 12-27 times lower activities compared to fish without additional clean feeding. After 18 weeks exposure, 137Cs levels in all the caged fish except for the control group were below the European permissible level (600 Bq kg-1) for consumption. In contrast, KFCF did not affect the intake of 90Sr in fish, although additional feed increased the growth rate and thereby accumulation in bone tissues. Hence, the use of clean feed containing KFCF is judged to be an effective and inexpensive countermeasure to reduce the 137Cs contamination of edible fish muscles.

Why is the multiple stressor concept of relevance to radioecology?

Purpose: A number of nuclear and radiological sources have contributed or are still contributing to the release of a series of stressors such as radionuclides in combination with trace metals and even organic chemicals. To assess the impact of mixed contamination, a limited number of stressors are usually evaluated one by one. We have therefore evaluated the benefit of using the multiple stressor concept focusing on key topics within radioecology such as the source term and deposition, ecosystem interactions and exposure, biological uptake and effects including adverse outcome pathways (AOPs), as a basis for assessing impact and risk. Materials and methods: We have extracted information from highly relevant scientific articles (e.g. Web of Science) describing multiple stressor exposure experiments where at least one stressor is ionizing radioactivity such as low dose gamma radiation or radionuclides (e.g. fission or activation products; uranium and daughter nuclides). In addition, experiences obtained during recent years at the author´s laboratories are included in the discussions, especially when it comes to speciation issues, combined effects, risk estimates, and AOPs. Results: The multiple stressor concept is not only related to the total concentration of a series of radionuclides and other stressors released from one source or from different sources, but also related to the presence of different physico-chemical forms of individual radionuclides. Due to interactions, the ecosystem transfer to biota is dynamic, and the application of toxicokinetics and toxicodynamics considerations seems most useful. Although there is a limited number of articles dealing with multiple stressors in which ionizing radiation is included as one of the stressors, deviation from concentration (dose) additive responses should probably be expected, especially under field conditions where additional abiotic as well as biotic interactions take place Conclusions: Following severe nuclear events, releases can be described as a massive multiple stressor exposure containing radionuclides combined with other stressors. Thus, advanced characterization technologies are needed for parameterization of the mixed stressor source term as input to ecosystem transport, dose, and impact models. To reduce uncertainties in assessments of these complex exposures, advanced technologies and computational efforts to link internal distributions to responses are also needed to understand how multiple stressor exposure could affect toxicokinetics and toxicodynamics, and why combined exposures could depart from additivity along the source-adverse outcome continuum.

The radiation exposure of fish in the period of the Techa river peak contamination.

Waterborne radioactive releases into the Techa River from the Mayak Production Association in Russia during 1949-1956 resulted in downstream contamination of the river ecosystem. The discharged liquid waste contained both short-lived isotopes (95Zr, 95Nb, 103,106Ru, 141,144Ce, 91Y, 89Sr and 140Ba with half-life from 3 days to 1.02 years) and the long-lived 90Sr and 137Cs (half-life - 28.79 y and 30.07 y, respectively). Even now, when two half-lives of 90Sr and 137Cs have passed, the contamination in the upper river region (about 70 km from the source of releases) is still relatively high. Current anthropogenic dose rates calculated for the fish of the Techa River depend on the distance along the stream and decrease from 150 to 3 µGy day-1. Radiation exposure of fish is expected to have been much higher at the time of the releases. The aim of the study was to evaluate the dose rates for the most common fish species of the river, viz., roach (Rutilus rutilus), perch (Perca fluviatilis) and pike (Esox lucius), in the period of peak contamination of the upper reaches of the Techa River from 1950 to 1951. To achieve this objective, calculation of both internal and external dose rates was performed. For dose rate calculation, the contamination of the river compartments was modeled, body-size dependent dose coefficients were evaluated, morphometric data were analyzed. Maximum dose rates were obtained for roach; minimum - for pike, it depends on fish lifestyle (time spent at the bottom). In the period before September 1950, fish of the upper reaches are assessed to have been exposed to dose rates exceeding the screening level equal to 240 µGy day-1. From September 1950 up to the end of 1952 the fish dose rates along the Techa River were found to be close to the UNSCEAR threshold equal to 9.6 × 103 µGy day-1 or even much more higher (up to 1.9 × 105 µGy day-1). Extremely high historical dose rates did not lead to the difference in fish size and fish growth rate currently observed in the Techa River and in the comparison waterbody (the Miass River). Discussion includes the description of radiation effects observed currently in the river fish. Today the effects observed in hematopoietic system may be the consequence of radiation exposure of fish over several generations. For example, long term dwelling of fish in the radioactively contaminated environment leads to their adaptation to chronic radiation exposure. At the same time, an increase their sensitivity and decrease their adaptive capacity to respond to other stress factors can be observed.

[Assessment of Erythropoiesis Status in Roach (Rutilus rutilus) of the Radioactively Contaminated Techa River].

At present volumetric activity of ß-emitting radionuclides in water at various locations of the Techa River ranges from 5 to 40-Bq/L; a specific activity of ß-emitting radionuclides in the bottom sediments at various locations ranges 10 Ito 106 Bq/kg dry weight. A significant increase of the erythroblast content in blood as compared to that in the roach from the reference watercourse (the Miass River) was observed during spawning in the spring. Due to this fact the number of erythrocytes was equal to that in the control animals under chronic radiation exposure at the dose rates of 0.9 and 16 µGy/day, and was insufficient at the dose rate of 108 gGy/day. During summer feeding no changes in the indexes of erythropoiesis in roach were observed under chronic radiation exposure at the dose rate of 0.9 µGy/day; the number of erythrocytes in the peripheral blood declines when the dose rates are 16 and 108 µGy/day. When performing a regression analysis, we revealed a dose-rate-dependent decrease in the absolute number of erythrocytes, normocytes, polychromatocytes, dividing and non-dividing erythroid cells in the peripheral blood of roach from the Techa River and an increase of a relative number of normochromatophylic erythrocytes.

Evaluation of distribution coefficients and concentration ratios of (90)Sr and (137)Cs in the Techa River and the Miass River.

Empirical data on the behavior of radionuclides in aquatic ecosystems are needed for radioecological modeling, which is commonly used for predicting transfer of radionuclides, estimating doses, and assessing possible adverse effects on species and communities. Preliminary studies of radioecological parameters including distribution coefficients and concentration ratios, for (90)Sr and (137)Cs were not in full agreement with the default values used in the ERICA Tool and the RESRAD BIOTA codes. The unique radiation situation in the Techa River, which was contaminated by long-lived radionuclides ((90)Sr and (137)Cs) in the middle of the last century allows improved knowledge about these parameters for river systems. Therefore, the study was focused on the evaluation of radioecological parameters (distribution coefficients and concentration ratios for (90)Sr and (137)Cs) for the Techa River and the Miass River, which is assumed as a comparison waterbody. To achieve the aim the current contamination of biotic and abiotic components of the river ecosystems was studied; distribution coefficients for (90)Sr and (137)Cs were calculated; concentration ratios of (90)Sr and (137)Cs for three fish species (roach, perch and pike), gastropods and filamentous algae were evaluated. Study results were then compared with default values available for use in the well-known computer codes ERICA Tool and RESRAD BIOTA (when site-specific data are not available). We show that the concentration ratios of (137)Cs in whole fish bodies depend on the predominant type of nutrition (carnivores and phytophagous). The results presented here are useful in the context of improving of tools for assessing concentrations of radionuclides in biota, which could rely on a wider range of ecosystem information compared with the process limited the current versions of ERICA and RESRAD codes. Further, the concentration ratios of (90)Sr are species-specific and strongly dependent on Ca(2+) concentration in water. The universal characteristic allows us to combine the data of fish caught in the water with different mineralization by multiplying the concentration of Ca(2+). The concentration ratios for fishes were well-fitted by Generalized Logistic Distribution function (GLD). In conclusion, the GLD can be used for probabilistic modeling of the concentration ratios in freshwater fishes to improve the confidence in the modeling results. This is important in the context of risk assessment and regulatory.

Aluminum exposure impacts brain plasticity and behavior in Atlantic salmon (Salmo salar).

Aluminum (Al) toxicity occurs frequently in natural aquatic ecosystems as a result of acid deposition and natural weathering processes. Detrimental effects of Al toxicity on aquatic organisms are well known and can have consequences for survival. Fish exposed to Al in low pH waters will experience physiological and neuroendocrine changes that disrupt homeostasis and alter behavior. To investigate the effects of Al exposure on both the brain and behavior, Atlantic salmon (Salmo salar) kept in water treated with Al (pH 5.7, 0.37±0.04 µmol 1(-1) Al) for 2 weeks were compared with fish kept in under control conditions (pH 6.7, <0.04 µmol 1(-1) Al). Fish exposed to Al and acidic conditions had increased Al accumulation in the gills and decreased gill Na(+), K(+)-ATPase activity, which impaired osmoregulatory capacity and caused physiological stress, indicated by elevated plasma cortisol and glucose levels. Here we show for the first time that exposure to Al in acidic conditions also impaired learning performance in a maze task. Al toxicity also reduced the expression of NeuroD1 transcript levels in the forebrain of exposed fish. As in mammals, these data show that exposure to chronic stress, such as acidified Al, can reduce neural plasticity during behavioral challenges in salmon, and may impair the ability to cope with new environments.

Effects of combined gamma-irradiation and metal (Al+Cd) exposures in Atlantic salmon (Salmo salar L.).

These experiments were designed to investigate transcriptional effects in Atlantic salmon (Salmo salar) after exposure in vivo to ionizing gamma radiation combined with subtoxic levels of aluminum (Al) and cadmium (Cd). Juvenile fish (35 g) in freshwater with or without Al and Cd (255 microg Al/L + 6 microg Cd/L) were exposed to a 75 mGy dose of gamma-irradiation, and induced responses were compared to those of controls. The transcriptional levels of eight genes encoding proteins known to respond to stress in fish were quantified in liver of fish exposed for 5 h to gamma radiation, to Al and Cd or to the combination of Al, Cd and gamma radiation. The studied genes were caspase 3B, caspase 6A, caspase 7, p53 (apoptosis), glutathione reductase (GR), phospholipid hydroperoxide glutathione peroxidase (GSH-Px), (oxidative stress), metallothionein (MT-A) (metal stress) and ubiquitin (Ubi) (protein degradation). The results showed that gamma-irradiation alone induced significant upregulation of caspase 6A, GR, GSH-Px, MT-A and Ubi compared to the control group, while 5 h exposure to Al+Cd alone did not induce any of the studied genes compared to the control. No significant upregulation of the series of investigated genes could be observed in fish exposed to gamma-irradiation in combination with Al+Cl. In conclusion, the results suggest that the presence of Al+Cd in the water counteracted the gamma-irradiation effect by modifying the transcription of genes encoding proteins involved in the defense mechanisms against free radicals in the cells.

Environmentally relevant mixed exposures to radiation and heavy metals induce measurable stress responses in Atlantic salmon.

These experiments were designed to identify stress effects in 3 key organs in Atlantic Salmon (Salmo salar, L.) after exposure in vivo to very low doses of radiation, and subtoxic levels of aluminum (Al) and cadmium (Cd) alone or in combination. Six fish per group were sacrificed after exposure and the anterior kidney, fin, and gill were dissected and sentfor assay of bystander signal production as a stress response end point. Radiation doses as low as 4 mGy delivered over 5 h, alone or in combination with Cd and/or Al, caused bystander signals to be produced in tissues harvested from in vivo exposed salmon. The effects vary among different organs and are not consistently additive or synergistic for a given treatment although gill cells do show high degrees of synergism between radiation and metal exposure. Data for individual fish did not suggest any systemic sensitivity to the stressors. Interestingly, the data for Cd suggest that lower toxicity is found when the metal is used in combination with radiation exposure. Expression of two proteins associated with survival responses (Bcl-2) or death responses (cmyc) after radiation was measured in the tissue cultures and showed a highly significant correlation with response outcome. The results, although complex, indicate that these stress signal responses may aid in the mechanistic investigation of mixed contaminant effects in fish exposed to metals and radiation.

Multiple stressor effects of radiation and metals in salmon (Salmo salar).

These experiments were designed to look at the cellular effects in key organs in Atlantic salmon (Salmo salar) after exposure in vivo to radiation and subtoxic levels of aluminum (Al) and cadmium (Cd), alone or in combination. Salmon (25g) were exposed to a single 0.5Gy dose of gamma-irradiation in water containing Cd, Al or Cd+Al. Three fish per group were sacrificed after 1h and the liver, pronephros, fin and gill of each was dissected. Small explants of each tissue were set up. After 2 days, the culture medium was harvested and filtered then placed on a reporter cell line for determination of stress signal activity (bystander effects). Radiation in combination with Cd and/or Al, caused bystander effects in tissues harvested from in vivo exposed salmon. The effects vary between different organs and are not consistently additive or synergistic for a given treatment. Tissue type appears to be critical. Liver cultures produce a toxic factor which is lethal to reporter cells, and therefore no liver data could be obtained. It is hoped that this stress signal response will prove to be a useful indicator of environmental stress in species inhabiting aquatic ecosystems.