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The shift towards a circular economy, where waste generation is minimized through waste re-use and the development of valorization strategies, is crucial for the establishment of a low carbon, sustainable, and resource-efficient economy. However, there is a lack of strategies for re-using and valorizing specific types of waste, particularly those containing naturally occurring radioactive materials (NORM), despite the prevalence of industrial activities that produce such waste due to their chemical and radiological hazards. Living organisms, including fungi, are valuable sources of bioactive compounds with various industrial applications. In this study, we assessed the growth and metabolic profile changes of three white rot fungi species in response to low concentrations of a uranium mine effluent containing NORM and metals to explore their potential for producing biotechnologically relevant bioactive compounds. The growth rate was assessed in three different culture media, with and without the uranium mine effluent (1% V/V)), and the metabolic profile was analyzed using FTIR-ATR spectroscopy. Results suggested an improvement in growth rates in media containing the uranium mine effluent, although not statistically significant. T. versicolor showed promise in terms of bioactive compound production. The production of droplets during growth experiments and significant metabolic changes, associated with the production of bioactive compounds like laccase, melanin, and oxalic acid, were observed in T. versicolor grown in mYEPDA with the uranium mine effluent. These findings present new research opportunities for utilizing waste to enhance the biotechnological production of industrially relevant bioactive compounds and promote the development of circular economy strategies for re-using and valorizing NORM-containing waste., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The bystander effect is commonly defined as the observation of effects in nonirradiated cells and tissues when the later are in contact with irradiated cells/ tissues. More recently the occurrence of bystander effect between organisms of the same species has been also demonstrated. Nevertheless, there is limited knowledge about this effect between soil dwelling organisms from different taxonomic groups, as well as in response to stressors other than ionizing radiation. Moreover, data reporting this phenomenon for soil invertebrates are scarce. The results herein presented contribute for the understanding of the impacts of cadmium and uranium in the DNA integrity of two terrestrial oligochaetes species (Eisenia fetida and Enchytraeus albidus). The evaluation was based on the quantification of the effects in the DNA integrity of the coelomocytes using the alkaline comet assay technique. This work reports the existence of bystander signaling from terrestrial earthworms to enchytraeids and from enchytraeids to earthworms when the organisms were exposed to Cd. These results reinforce that the bystander effect seems to be related with the genotoxic activity of stressors, and not exclusive of radiotoxic contaminants. Further, the bystander effect occurs between different species and under real environmental conditions, even in complex matrices, as the soil., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Uranium mining sites produce residues rich in metals and radionuclides, that may contaminate all environmental matrices, exposing human and non-human biota to low doses of ionizing radiation (LDIR) and to the chemical toxicity of several metals. To date, experimental and radio-epidemiological studies do not provide conclusive evidence of LDIR induced cancer. However, co-exposures (LDIR plus other contaminants), may increase the risks. To determine the potential for genotoxic effects in human cells induced by the exposure to LDIR plus metals, HEPG2 cells were exposed to different concentrations of a uranium mine effluent for 96 h. DNA damage was evaluated using the comet assay and changes in the expression of tumor suppressor and oncogenes were determined using qPCR. Results show that effluent concentrations higher than 5%, induce significant DNA damage. Also, a significant under-expression of ATM and TP53 genes and a significant overexpression of GADD45a gene was observed. Results show that the exposure to complex mixtures cannot be disregarded, as effects were detected at very low doses. This study highlights the need for further studies to clarify the risks of exposure to LDIR along with other stressors, to fully review the IR exposure risk limits established for human and non-human biota., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Assessing the impact of uranium mining industry on aquatic ecosystems near mining areas is critical to ensure the long-term health and sustainability of ecosystem services. As so, a transgenerational study with Daphnia magna has been conducted to perceive to what extent intermittent discharges of uranium mine effluents into watercourses may impact the DNA integrity and life history traits of cladocerans. Organisms were exposed for 48 h to a 2% dilution of an uranium mine effluent (UME) and to a corresponding dose of waterborne uranium (WU) that, according to our preliminary studies, induces significant DNA damage in daphnids. After exposure, organisms were transferred to clean medium, where three successive generations were monitored for genotoxicity and other effects at the individual and population level. Despite some differences between WU and UME data, our results revealed that the negative impacts of the short-term exposure gradually disappeared after placing the organisms in clean medium. These results suggest that, under intermittent stress, daphnids are able to recover, since after the 3 rd brood release, DNA damage (measured as DNA strand breaks) is no longer observed and has no significant impact on the detectable life traits of offspring. Although our results indicate that populations of D. magna are not affected by intermittent and highly diluted discharges from uranium mining, aquatic systems under this kind of pressure should not be seen as hazardous-free. Future studies in this field are recommended and these should consider radionuclides in the water column, their accumulation in the sediments and also multiple life stages., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The induction of RIBE (Radiation Induced Bystander Effect) is a non-target effect of low radiation doses that has already been verified at an inter-organismic level in fish and small mammals. Although the theoretical impact in the field of environmental risk assessment (ERA) is possible, there is a gap of knowledge regarding this phenomenon in invertebrate groups and following environmentally relevant exposures. To understand if RIBE should be considered for ERA of radionuclide-rich wastewaters, we exposed Daphnia magna (<24 h and 5d old) to a 2% diluted uranium mine effluent for 48 h, and to a matching dose of waterborne uranium (55.3 μg L -1 ). Then the exposed organisms were placed (24 and 48 h) in a clean medium together with non-exposed neonates. The DNA damage observed for the non-exposed organisms was statistically significant after the 24 h cohabitation for both uranium (neonates p = 0.002; 5 d-old daphnids p = <0.001) and uranium mine effluent exposure (only for neonates p = 0.042). After 48 h cohabitation significant results were obtained only for uranium exposure (neonates p = 0.017; 5 d-old daphnids p = 0.013). Although there may be some variability associated to age and exposure duration, the significant DNA damage detected in non-exposed organisms clearly reveals the occurrence of RIBE in D. magna. The data obtained and here presented are a valuable contribution for the discussion about the relevance of RIBE for environmental risk assessment., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Active and abandoned uranium mining sites often create environmentally problematic situations, since they cause the contamination of all environmental matrices (air, soil and water) with stable metals and radionuclides. Due to their cytotoxic, genotoxic and teratogenic properties, the exposure to these contaminants may cause several harmful effects in living organisms. The Fish Embryo Acute Toxicity Test (FET) test was employed to evaluate the genotoxic and teratogenic potential of mine liquid effluents and sludge elutriates from a deactivated uranium mine. The aims were: a) to determine the risk of discharge of such wastes in the environment; b) the effectiveness of the chemical treatment applied to the uranium mine water, which is a standard procedure generally applied to liquid effluents from uranium mines and mills, to reduce its toxicological potential; c) the suitability of the FET test for the evaluation the toxicity of such wastes and the added value of including the evaluation of genotoxicity. Results showed that through the FET test it was possible to determine that both elutriates and effluents are genotoxic and also that the mine effluent is teratogenic at low concentrations. Additionally, liquid effluents and sludge elutriates affect other parameters namely, growth and hatching and that water pH alone played an important role in the hatching process. The inclusion of genotoxicity evaluation in the FET test was crucial to prevent the underestimation of the risks posed by some of the tested effluents/elutriates. Finally, it was possible to conclude that care should be taken when using benchmark values calculated for specific stressors to evaluate the risk posed by uranium mining wastes to freshwater ecosystems, due to their chemical complexity., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Concerns about the impacts on public health and on the natural environment have been raised regarding the full range of operational activities related to uranium mining and the rest of the nuclear fuel cycle (including nuclear accidents), nuclear tests and depleted uranium from military ammunitions. However, the environmental impacts of such activities, as well as their ecotoxicological/toxicological profile, are still poorly studied. Herein, it is discussed if organisms can be used as bioindicators of human health effects, posed by lifetime exposure to radioactively contaminated areas. To do so, information was gathered from several studies performed on vertebrates, invertebrate species and humans, living in these contaminated areas. The retrieved information was compared, to determine which are the most used bioindicators and biomarkers and also the similarities between human and non-human biota responses. The data evaluated are used to support the proposal for an early warning scheme, based on bioindicator species and on the most sensitive and commonly shared biomarkers, to perform a screening evaluation of radioactively contaminated sites. This scheme could be used to support decision-making for a deeper evaluation of risks to human health, making it possible to screen a large number of areas, without disturbing and alarming local populations., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Environmental exposure to uranium and its daughter radionuclides, has been linked to several negative effects such as those related with important physiological processes, like hematopoiesis, and may also be associated with genotoxicity effects. Herein, genotoxic effects, immunotoxicity, trace elements and C reactive protein (CRP) analyses, were performed in peripheral blood samples collected from individuals of a population living near a deactivated uranium mine. C reactive protein analysis was performed to exclude candidates with active inflammatory processes from further evaluations. DNA damage and immunotoxicity (immunophenotyping and immune cell counts) were evaluated by comet assay and flow cytometry, respectively. Significant DNA damage was observed in the peripheral blood samples from volunteers living in the Cunha Baixa village. A significant decrease of NK and T lymphocytes counts were observed in the individuals from the Cunha Baixa village, when compared with individuals from the reference site. Uranium and manganese levels were significantly higher in the Cunha Baixa village inhabitants. On the other hand, zinc levels were significantly lower in those individuals when compared with the volunteers from the control village. Results suggest that inhabitants from Cunha Baixa have a higher risk of suffering from serious diseases such as cancer, since high DNA damages were observed in peripheral blood leukocytes and also decreased levels of NK and T cells, which play an essential role in the defense against tumor growth., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

The effects of the exposure of earthworms (Eisenia andrei) to contaminated soil from an abandoned uranium mine, were assessed through gene expression profile evaluation by Suppression Subtractive Hybridization (SSH). Organisms were exposed in situ for 56 days, in containers placed both in a contaminated and in a non-contaminated site (reference). Organisms were sampled after 14 and 56 days of exposure. Results showed that the main physiological functions affected by the exposure to metals and radionuclides were: metabolism, oxireductase activity, redox homeostasis and response to chemical stimulus and stress. The relative expression of NADH dehydrogenase subunit 1 and elongation factor 1 alpha was also affected, since the genes encoding these enzymes were significantly up and down-regulated, after 14 and 56 days of exposure, respectively. Also, an EST with homology for SET oncogene was found to be up-regulated. To the best of our knowledge, this is the first time that this gene was identified in earthworms and thus, further studies are required, to clarify its involvement in the toxicity of metals and radionuclides. Considering the results herein presented, gene expression profiling proved to be a very useful tool to detect earthworms underlying responses to metals and radionuclides exposure, pointing out for the detection and development of potential new biomarkers., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Genotoxic effects caused by the exposure to wastes containing metals and radionuclides were investigated in the European wood mice (Apodemus sylvaticus). The animals were captured in the surroundings of an abandoned uranium mining site. DNA damage was assessed by comet assay; gene expression and single nucleotide polymorphisms (SNPs) were assessed, respectively, by Real-Time PCR and melt curve analysis. The bioaccumulation of metals in the liver, kidney and bones was also determined to help clarify cause-effect relationships. Results confirmed the bioaccumulation of cadmium and uranium in organisms exposed to uranium mining wastes. P53 gene was found to be significantly up-regulated in the liver of those organisms and SNPs in the Rb gene were also detected in the kidney. Our results showed that uranium mining wastes caused serious DNA damage resulting in genomic instability, disclosed by the significant increase in DNA strand breaks and P53 gene expression disturbance. These effects can have severe consequences, since they may contribute for the emergence of serious genetic diseases. The fact that mice are often used as bioindicator species for the evaluation of risks of environmental exposure to humans, raises concerns on the risks for human populations living near uranium mining areas., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Earthworms were exposed for 56 days to a contaminated soil from an abandoned uranium mine and to the natural reference soil LUFA 2.2. The exposure occurred in situ: the containers with contaminated soil were placed near the mine pit; the containers with reference soil were placed in a reference site. For the assessment of metals bioaccumulation, DNA damages, cell-to-cell variation in DNA content, Median Fluorescence Intensity (MFI), coelomocytes frequency and proliferation, organisms were sampled after 0, 1, 2, 7, 14 and 56 days of exposure. For the assessment of radionuclides bioaccumulation, animals were sampled after 0, 14 and 56 days of exposure. As for growth, organisms were sampled after 0, 14, 28 and 56 days of exposure. The reproduction assay was performed according to the OECD (2004) guideline. DNA damages were assessed by comet assay and flow cytometry was used to determine cell-to-cell variation in DNA content, Median Fluorescence Intensity (MFI), coelomocytes frequency and proliferation. Results have shown a myriad of effects in the organisms exposed to the contaminated soil, namely: the inhibition of reproduction, growth reduction, DNA damages, cytotoxicity, changes in eleocytes fluorescence intensity, coelomocytes proliferation and bioaccumulation of metals and radionuclides. Our results showed that the evaluation of genotoxicity and cytotoxicity endpoints, along with other parameters at an individual level in standard reproduction assays conducted in situ, are important to improve the risk assessment process of areas contaminated with uranium and other radioactive mining wastes., (Copyright © 2011 Elsevier Inc. All rights reserved.)