The effect of bacterial growth phase and culture concentration on U(VI) removal from aqueous solution

Bacteria play a key role in controlling the mobility of contaminants, such as uranium (U), in the environment. Uranium could be sourced from disposed radioactive waste, derived either from surface disposal trenches for Low Level Waste (LLW) that, because of the waste type and disposal concept, would typically present acidic conditions or from the geological disposal of LLW or Intermediate Level Waste (ILW) that, because of the waste type and the disposal concept, would typically present alkaline conditions. In disposed radioactive waste, there could be variable amounts of cellulosic material. Bacterial cells may be living in a range of different growth phases, depending on the growth conditions and nutrients available at the time any waste-derived U migrated to the cells. A key knowledge gap to date has been the lack of a mechanistic understanding of how bacterial growth phases (exponential, stationary, and death phase) affect the ability of bacteria to remove U(VI) from solution. To address this, we first characterised the cells using potentiometric titrations to detect any differences in proton binding to proton active sites on Pseudomonas putida cells at each growth phase under aerobic conditions, or under anaerobic conditions favourable to U(IV) reoxidation. We then conducted batch U(VI) removal experiments with bacteria at each phase suspended in 1 and 10 ppm U aqueous solutions with the pH adjusted from 2 to 12 as well as with culture concentrations from 0.01 to 10 g/L, to identify the minimal concentration of bacteria in solution necessary to affect U removal. We found that, in death phase, P. putida cells exhibited double the concentration of proton active sites than bacteria grown to exponential and stationary phase. However, we did not see a difference in the extent of U(VI) removal, from a 10 ppm U solution, between the different growth phases as a function of pH (2 to 12). Culture concentration affected U removal between pH 2–8, where U removal decreased with a decreasing concentration of cells in solution. When the pH was 10–12, ≤55% of U precipitated abiotically. The presence of bacteria in solution (0.01–10 g/L), regardless of growth phase, increased the precipitation of U from ≤55% up to 70–90%, accumulating inside the cells and on the cell walls as ~0.2 μm uranyl phosphate precipitates. These precipitates were also found at low pH with the exception of cells at exponential growth phase. This study demonstrates that growth phase affects the proton-active site concentration but not the extent of U bound to P. putida cells and that growth phase dictates the form of U removed from solution. Since the pH of trench-disposed LLW is controlled by the degradation of cellulosic waste, leading to acidic conditions (pH 4–6), bacterial concentrations would be expected to highly affect the extent of U removed from solution. The cement in grouted ILW and LLW, for geologic disposal, will allow for the development of extremely high pH values in solution (pH 9–13), where even the smallest concentrations of bacteria were able to significantly increase the removal of U from solution under aerobic conditions, or under anaerobic conditions favourable to U(IV) reoxidation

Cell surface acid-base properties of the cyanobacterium Synechococcus: Influences of nitrogen source, growth phase and N:P ratios

The distribution of many trace metals in the oceans is controlled by biological uptake. Recently, Liu et al. (2015) demonstrated the propensity for a marine cyanobacterium to adsorb cadmium from seawater, suggesting that cell surface reactivity might also play an important role in the cycling of metals in the oceans. However, it remains unclear how variations in cyanobacterial growth rates and nutrient supply might affect the chemical properties of their cellular surfaces. In this study we used potentiometric titrations and Fourier Transform Infrared (FT-IR) spectrometry to profile the key metabolic changes and surface chemical responses of a Synechococcus strain, PCC 7002, during different growth regimes. This included testing various nitrogen (N) to phosphorous (P) ratios (both nitrogen and phosphorous dependent), nitrogen sources (nitrate, ammonium and urea) and growth stages (exponential, stationary, and death phase). FT-IR spectroscopy showed that varying the growth substrates on which Synechococcus cells were cultured resulted in differences in either the type or abundance of cellular exudates produced or a change in the cell wall components. Potentiometric titration data were modeled using three distinct proton binding sites, with resulting pKa values for cells of the various growth conditions in the ranges of 4.96-5.51 (pKa1), 6.67-7.42 (pKa2) and 8.13-9.95 (pKa3). According to previous spectroscopic studies, these pKa ranges are consistent with carboxyl, phosphoryl, and amine groups, respectively. Comparisons between the titration data (for the cell surface) and FT-IR spectra (for the average cellular changes) generally indicate (1) that the nitrogen source is a greater determinant of ligand concentration than growth phase, and (2) that phosphorus limitation has a greater impact on Synechococcus cellular and extracellular properties than does nitrogen limitation. Taken together, these techniques indicate that nutritional quality during cell growth can noticeably influence the expression of cell surface ligands and their measurable densities. Given that cell surface charge ultimately affects metal adsorption, our results suggest that the cycling of metals by Synechococcus cells in the oceans may vary regionally

Fate of trace metals in anaerobic digestion

© Springer International Publishing Switzerland 2015. A challenging, and largely uncharted, area of research in the field of anaerobic digestion science and technology is in understanding the roles of trace metals in enabling biogas production. This is a major knowledge gap and a multifaceted problem involving metal chemistry; physical interactions of metal and solids; microbiology; and technology optimization. Moreover, the fate of trace metals, and the chemical speciation and transport of trace metals in environments— often agricultural lands receiving discharge waters from anaerobic digestion processes— simultaneously represents challenges for environmental protection and opportunities to close process loops in anaerobic digestion.The authors acknowledge funding within the framework of the COST Action 1302 (‘European Network on Ecological Roles of Trace Metals in Anaerobic Biotechnologies’). GC is supported by a European Research Council Starting Grant (‘3C-BIOTECH; No. 261330).Peer Reviewe