Continuous cultivation of the lithoautotrophic nitrate-reducing Fe(II)-oxidizing culture KS in a chemostat bioreactor

Laboratory-based studies on microbial Fe(II) oxidation are commonly performed over just a few weeks in small volumes with high substrate concentrations, resulting in geochemical gradients and volumetric effects caused by sampling. We used a chemostat to enable uninterrupted supply of medium, and investigated autotrophic growth of the nitrate-reducing Fe(II)-oxidizing culture KS for 24 days. We analysed Fe- and N-speciation, cell-mineral associations, and the identity of minerals. Results were compared to different batch systems (50 and 700 ml – static/shaken). The Fe(II) oxidation rate was highest in the chemostat with 7.57 mM Fe(II) d-1, while the extent was similar (averaged 92% of all Fe(II)). Short-range ordered Fe(III) phases, presumably ferrihydrite, precipitated and later goethite was detected in the chemostat. 1 mM solid phase Fe(II) remained in the chemostat, up to 15 µM of reactive nitrite was measured, and 42% of visualized cells were partially or completely mineral-encrusted, likely caused by abiotic oxidation of Fe(II) by nitrite. Despite (partial) encrustation, cells were still viable. Our results show that even with similar oxidation rates as in batch cultures, cultivating Fe(II)-oxidizing microorganisms under continuous conditions reveals mechanistic insights on the role of reactive intermediates for Fe(II) oxidation, mineral formation and cell-mineral interactions.

Production of Hydroxyl Radicals From Oxygenation of Simulated AMD In Presence of Extracellular Polymers Substances

While the reaction mechanisms Fe(II) abiotic oxidation produce ·OH by CaCO3-induced in AMD are well-documented, little is known about the influence of extracellular polymeric substances (EPS) secreted by microorganisms on Fe(II) oxidation in AMD. Given the recent finding, this study experimently measured the cumulative concentrations of ·OH produced from oxygenation of simulated AMD in the presence of EPS. Results of this study show that the cumulative ·OH increased from 56.75 to 158.70 μM within 24 h at pH 3 with the increase in EPS concentration from 0 to 12 mg/L. An appropriate pH (about 6) and EPS (6 mg/L) concentration were required for the moderate rate of Fe(II) oxidation, corresponding to the maximum production of ·OH. The presence of EPS enhanced the ·OH production from Fe(II) oxidation in simulated AMD under acid conditions. In the presence of EPS, ·OH production is attributed mainly the complexation of Fe(II) with EPS, of which is rich of carboxyl and hydroxyl groups. Besides, the yield of ·OH increased remarkably with the addition of Fe3+. It is most likely that EPS can contribute to reduce Fe(Ⅲ) to Fe(II), which is beneficial to the production of ·OH. The findings reveal from this study supplement the fundamental of ·OH production from Fe(II) oxidation by microorganisms in natural AMD.

Mn oxide formation by phototrophs: Spatial and temporal patterns, with evidence of an enzymatic superoxide-mediated pathway

Manganese (Mn) oxide minerals influence the availability of organic carbon, nutrients and metals in the environment. Oxidation of Mn(II) to Mn(III/IV) oxides is largely promoted by the direct and indirect activity of microorganisms. Studies of biogenic Mn(II) oxidation have focused on bacteria and fungi, with phototrophic organisms (phototrophs) being generally overlooked. Here, we isolated phototrophs from Mn removal beds in Pennsylvania, USA, including fourteen Chlorophyta (green algae), three Bacillariophyta (diatoms) and one cyanobacterium, all of which consistently formed Mn(III/IV) oxides. Isolates produced cell-specific oxides (coating some cells but not others), diffuse biofilm oxides, and internal diatom-specific Mn-rich nodules. Phototrophic Mn(II) oxidation had been previously attributed to abiotic oxidation mediated by photosynthesis-driven pH increases, but we found a decoupling of Mn oxide formation and pH alteration in several cases. Furthermore, cell-free filtrates of some isolates produced Mn oxides at specific time points, but this activity was not induced by Mn(II). Manganese oxide formation in cell-free filtrates occurred via reaction with the oxygen radical superoxide produced by soluble extracellular proteins. Given the known widespread ability of phototrophs to produce superoxide, the contribution of phototrophs to Mn(II) oxidation in the environment may be greater and more nuanced than previously thought.