Soil microbial community structure and functionality changes in response to long-term metal and radionuclide pollution.

Microbial communities are essential for a healthy soil ecosystem. Metals and radionuclides can exert a persistent pressure on the soil microbial community. However, little is known on the effect of long-term co-contamination of metals and radionuclides on the microbial community structure and functionality. We investigated the impact of historical discharges of the phosphate and nuclear industry on the microbial community in the Grote Nete river basin in Belgium. Eight locations were sampled along a transect to the river edge and one location further in the field. Chemical analysis demonstrated a metal and radionuclide contamination gradient and revealed a distinct clustering of the locations based on all metadata. Moreover, a relation between the chemical parameters and the bacterial community structure was demonstrated. Although no difference in biomass was observed between locations, cultivation-dependent experiments showed that communities from contaminated locations survived better on singular metals than communities from control locations. Furthermore, nitrification, a key soil ecosystem process seemed affected in contaminated locations when combining metadata with microbial profiling. These results indicate that long-term metal and radionuclide pollution impacts the microbial community structure and functionality and provides important fundamental insights into microbial community dynamics in co-metal-radionuclide contaminated sites.

Effects of environmental parameters on Lemna minor growth: An integrated experimental and modelling approach.

Pollution of surface waters is a worldwide problem for people and wildlife. Remediation and phytoremediation approaches can offer a solution to deal with specific scenarios. Lemna minor, commonly known as duckweed, can absorb and accumulate pollutants in its biomass. To evaluate if L. minor could be applied for phytoremediation purposes, it is necessary to further investigate its remediation capability and to identify which parameters affect the remediation process. Such a model must include both plant growth and pollutant exchange. A remediation model based on a robust experimental study can help to evaluate L. minor as a proper remediation strategy and to predict the outcome of a L. minor based remediation system. To set up this model, this paper focusses on a detailed experimental study and a comprehensive mathematical modelling approach to represent L. minor growth as a function of biomass, temperature, light irradiation and variable nutrient concentrations. The influence of environmental conditions on L. minor growth was studied, by composing 7 days growth curves. Plants were grown under predefined environmental conditions (25°C, 14h photoperiod, 220 µmol m-2 s-1 light intensity and a modified Hoagland solution with 23.94 mg N L-1 and 3.10 mg P L-1 (N:P ratio of 7.73)) as standard for all experiments. The influence of different temperatures (6, 10, 15, 20, 25, 30 and 35°C), light intensities (63, 118, 170, 220 and 262 µmol m-2 s-1), photoperiods (12h and 14h) and N:P ratios (1.18, 3.36, 7.73 and 29.57) were tested in the model. As a result, a growth model was optimised using separate datasets for temperature, light intensity, photoperiod and nutrients and validated by further integrated testing. The growth model is a stable platform for application in phytoremediation of radionuclides in contaminated water, to be extended in future studies with information of pollutant uptake, pollutant-nutrient interactions and transfer to the biomass.

Rhizophagus irregularis MUCL 41833 can colonize and improve P uptake of Plantago lanceolata after exposure to ionizing gamma radiation in root organ culture.

Long-lived radionuclides such as (90)Sr and (137)Cs can be naturally or accidentally deposited in the upper soil layers where they emit ß/γ radiation. Previous studies have shown that arbuscular mycorrhizal fungi (AMF) can accumulate and transfer radionuclides from soil to plant, but there have been no studies on the direct impact of ionizing radiation on AMF. In this study, root organ cultures of the AMF Rhizophagus irregularis MUCL 41833 were exposed to 15.37, 30.35, and 113.03 Gy gamma radiation from a (137)Cs source. Exposed spores were subsequently inoculated to Plantago lanceolata seedlings in pots, and root colonization and P uptake evaluated. P. lanceolata seedlings inoculated with non-irradiated AMF spores or with spores irradiated with up to 30.35 Gy gamma radiation had similar levels of root colonization. Spores irradiated with 113.03 Gy gamma radiation failed to colonize P. lanceolata roots. P content of plants inoculated with non-irradiated spores or of plants inoculated with spores irradiated with up to 30.35 Gy gamma radiation was higher than in non-mycorrhizal plants or plants inoculated with spores irradiated with 113.03 Gy gamma radiation. These results demonstrate that spores of R. irregularis MUCL 41833 are tolerant to chronic ionizing radiation at high doses.

Highly weathered mineral soils have highest transfer risk of radiocaesium contamination after a nuclear accident: A global soil-plant study.

Accidental release of radiocaesium (137Cs) from nuclear power plants may result in long-term contamination of environmental and food production systems. Assessment of food chain contamination with 137Cs relies on 137Cs soil-to-plant transfer data and models mainly available for regions affected by the Chornobyl and Fukushima accidents. Similar data and models are lacking for other regions. Such information is needed given the global expansion of nuclear energy. We collected 38 soils worldwide of contrasting parent materials and weathering stages. The soils were spiked with 137Cs and sown with ryegrass in greenhouse conditions. The 137Cs grass-soil concentration ratio varied four orders of magnitude among soils. It was highest in Ferralsols due to the low 137Cs interception potential of kaolinite clay and the low exchangeable potassium in these soils. Our results demonstrate, for the first time, the high plant uptake of 137Cs in tropical soils. The most recent 137Cs transfer model, mainly calibrated to temperate soils dominated by weathered micas, poorly predicts the underlying processes in tropical soils but, due to compensatory effect, still reasonably well predicts 137Cs bioavailability across all soils (R2 = 0.8 on a log-log scale).

Differentiation between chemo- and radiotoxicity of 137Cs and 60Co on Lemna minor.

The uptake and effects of stable Cs and Co on L.minor were extensively studied, together with the effects of gamma radiation using a 137Cs or 60Co source. Innovative is that we combined external irradiation (from 137Cs or 60Co sources) with the direct uptake of certain amounts of stable Cs or Co to simulate the impact of the same mass of a radioisotope compared with that of the stable element. Such approach allows to differentiate between chemo- and radiotoxicity of 137Cs or 60Co, permitting to study the 137Cs and 60Co uptake by L. minor without using high concentrations of these elements in solution. Our results indicate that radiotoxicity of both 137Cs and 60Co has a greater importance compared to their chemotoxicity. This was also supported by the independent action and concentration addition concepts. Both concepts resulted in a good prediction of the dose-response curve of the combination exposure. The maximal removal of 137Cs or 60Co per gram dry matter of L. minor was lower compared with the removal of the corresponding stable isotope. The toxicity of 60Co was higher compared to 137Cs based on EC50 values and uptake data. With respect to the effects on photosynthetic pigments, starch and soluble sugars contents, only starch increased in a concentration- and dose-dependent manner.

pH and microbial community determine the denitrifying activity in the presence of nitrate-containing radioactive waste.

An important fraction of the currently stored volume of long-lived intermediate-level radioactive waste in Belgium contains large amounts of NaNO3 homogeneously dispersed in a hard bituminous matrix. Geological disposal of this waste form in a water-saturated sedimentary formation such as Boom Clay will result in the leaching of high concentrations of NaNO3, which could cause a geochemical perturbation of the surrounding clay, possibly affecting some of the favorable characteristics of the host formation. In addition, hyper-alkaline conditions are expected for thousands of years, imposed by the cementitious materials used as backfill material. Microbial nitrate reduction is a well-known process and can result in the accumulation of nitrite or nitrogenous gases. This could lead to the oxidation of redox-active Boom Clay components, which could (locally) decrease the reducing capacity of the clay formation. Here, we compared nitrate reduction processes between two microbial communities at different pH related to a geological repository environment and in the presence of a nitrate-containing waste simulate during 1 year in batch experiments. We showed that the microbial community from in Boom Clay borehole water was able to carry out nitrate reduction in the presence of acetate at pH 10.5, although the maximum rate of 1.3 ± 0.2 mM NO3 -/day was much lower compared to that observed at pH 9 (2.9 mM NO3 -/day). However, microbial activity at pH 10.5 was likely limited by a phosphate shortage. This study further confirmed that the Harpur Hill sediment harbors a microbial community adapted to high pH conditions. It reduced twice as much nitrate at pH 10.5 compared to pH 9 and the maximum nitrate reduction rate was higher at pH 10.5 compared to that at pH 9, i.e., 3.4 ± 0.8 mM NO3 -/day versus 2.2 ± 0.4 mM NO3 -/day. Both communities were able to form biofilms on non-radioactive Eurobitum. However, for both microbial communities, pH 12.5 seems to be a limiting condition for microbial activity as no nitrate reduction nor biofilm was observed. Nevertheless, pH alone is not sufficient to eliminate microbial presence, but it can induce a significant shift in the microbial community composition and reduce its nitrate reducing activity. Furthermore, at the interface between the cementitious disposal gallery and the clay host rock, the pH will not be sufficiently high to inhibit microbial nitrate reduction.

Distribution and behaviour of naturally occurring radionuclides within a Scots pine forest grown on a CaF2 waste deposit related to the Belgian phosphate industry.

The distribution and behaviour of naturally occurring radionuclides within a vegetated part of a CaF2 sludge heap from the Belgian phosphate industry was studied. A Scots pine forest plot was selected as study area. Trees were approximately 20 years old and showed a disturbed health state. Seasonal sampling campaigns of soil, roots, wood, inner and outer bark, needles and twigs gave insight on 238U, 226Ra, 210Pb and 210Po transfer and distribution between pine tree compartments. Soil samples were analysed for their texture, total organic and inorganic carbon, field capacity, pH and radionuclide content. Solid-liquid distribution coefficients (Kd) were experimentally determined for 238U, 226Ra (using Ba as analogue) and 210Pb based on adsorption-desorption batch tests. Results indicated higher 238U, 232Th, 226Ra, 210Pb and 210Po activity concentrations in the deeper soil layers while the first 20 cm contained less radionuclides but had a higher level of organic carbon. Additionally, results indicated no seasonal changes in the 238U:226Ra ratio in the soil while the 226Ra:210Pb ratio was significantly higher in spring compared to winter in the 20-60 cm soil layer. Pine tree roots served as natural translocation barrier for all radionuclides with high retention in the roots and low translocation to the above ground tree compartments. When considering the above ground compartments, 210Pb and 210Po were mostly present in the bark, needles and twigs. Furthermore, 238U and its progeny were highly accumulated in mosses. These results allowed us to establish more realistic soil-to-plant transfer factors. In addition, experimentally mimicking pore water acidification in the root zone resulted in lower 238U and 210Pb Kd values compared to using a standard CaCl2 solution. This study provides an integrated radioecological picture of knowledge and site specific data needed to study the long-term influence of vegetation on radionuclide dispersion in forest ecosystems.

Exposure to ionizing radiation affects the growth of ectomycorrhizal fungi and induces increased melanin production and increased capacities of reactive oxygen species scavenging enzymes.

Ectomycorrhizal (EM) fungi form symbioses with dominant tree families in boreal, temperate and tropical ecosystems and are important drivers of ecosystem function. EM fungal hyphae extend over a large area making them susceptible to enhanced radiation levels from naturally occurring or anthropogenically originating radioisotopes in the rhizosphere. In this study, the in-vitro effects of ionizing radiation on the growth and biomass of EM fungi Suillus luteus, S. bovinus and Rhizopogon luteolus were investigated. EM fungal cultures were exposed to gamma radiation from a 137Cs source for 137 h in darkness at 21 °C at dose rates of 404, 108.5 and 54.9 mGy h-1 resulting in total absorbed doses of 55.21, 14.82 and 7.50 Gy respectively. Cultures grown in the dark at 21 °C but not exposed to the 137Cs source served as the control. Our results show that EM fungi vary in their sensitivity to ionizing radiation. EM fungi used in this study produced melanin and reactive oxygen species scavenging enzymes such as catalase and superoxide dismutase as a response to ionizing radiation.

Arabidopsis plants exposed to gamma radiation in two successive generations show a different oxidative stress response.

When terrestrial environments get contaminated with long-lived gamma emitting radionuclides, plants that grow in these contaminated areas are exposed to gamma radiation during consecutive generations. Therefore it is important to evaluate the gamma induced stress response in plants in and between generations. The objective of this research is to reveal differences at the level of the antioxidative stress response between generations with a different radiation history. An experiment was conducted in which 7-days old Arabidopsis thaliana plants were exposed for 14 days to four different gamma dose rates: 22 mGy/h, 38 mGy/h, 86 mGy/h and 457 mGy/h. Two different plant groups were used: plants that were not exposed to gamma radiation before (P0) and plants that received the aforementioned gamma treatment during their previous generation (S1). Growth, the concentration of the antioxidants ascorbate and glutathione, a number of antioxidative enzyme activities and their gene transcript levels were analysed. A dose-rate dependent induction was seen for catalase (CAT) and guaiacol peroxidase (GPX) in the roots and for syringaldazine peroxidase (SPX) in the shoots. Differences between the two generations were observed for CAT and GPX in the roots, where a significantly higher activity of these reactive oxygen species (ROS) detoxifying enzymes was observed in the S1 generation. For SPX in the shoots, a dose dependent upregulation was observed in the P0 generation. However, high SPX activities were present for all doses in the S1 generation. These differences in enzyme activity between generations for SPX and GPX and the involvement of these enzymes in cell wall biosynthesis, suggest an important role for cell wall strengthening in the response to gamma irradiation.