Quantitative NME microscopy of iron transport in methanogenic aggregates

Transport of micronutrients (iron, cobalt, nickel, etc.) within biofilms matrixes such as methanogenic granules is of high importance, because these are either essential or toxic for the microorganisms living inside the biofilm. The present study demonstrates quantitative measurements of metal transport inside these biofilms using T1 weighted 3D RARE. It is shown that iron(II)-EDTA diffusion within the granule is independent of direction or the inner structure of the granules. Assuming position dependence of the spin-lattice relaxivity, Fick’s law for diffusion in a sphere can be applied to simulate the diffusion within the methanogenic granules under investigation. A relatively low diffusion coefficient of 2.5*10-11 m2·s-1 was obtained for iron diffusion within the methanogenic granul

Role of nickel in high rate methanol degradation in anaerobic granular sludge bioreactors

The effect of nickel deprivation from the influent of a mesophilic (30°C) methanol fed upflow anaerobic sludge bed (UASB) reactor was investigated by coupling the reactor performance to the evolution of the Methanosarcina population of the bioreactor sludge. The reactor was operated at pH 7.0 and an organic loading rate (OLR) of 5–15 g COD l−1 day−1 for 191 days. A clear limitation of the specific methanogenic activity (SMA) on methanol due to the absence of nickel was observed after 129 days of bioreactor operation: the SMA of the sludge in medium with the complete trace metal solution except nickel amounted to 1.164 (±0.167) g CH4-COD g VSS−1 day−1 compared to 2.027 (±0.111) g CH4-COD g VSS−1 day−1 in a medium with the complete (including nickel) trace metal solution. The methanol removal efficiency during these 129 days was 99%, no volatile fatty acid (VFA) accumulation was observed and the size of the Methanosarcina population increased compared to the seed sludge. Continuation of the UASB reactor operation with the nickel limited sludge lead to incomplete methanol removal, and thus methanol accumulation in the reactor effluent from day 142 onwards. This methanol accumulation subsequently induced an increase of the acetogenic activity in the UASB reactor on day 160. On day 165, 77% of the methanol fed to the system was converted to acetate and the Methanosarcina population size had substantially decreased. Inclusion of 0.5 μM Ni (dosed as NiCl2) to the influent from day 165 onwards lead to the recovery of the methanol removal efficiency to 99% without VFA accumulation within 2 days of bioreactor operation

Developments and constraints in fermentative hydrogen production

Fermentative hydrogen production is a novel aspect of anaerobic digestion. The main advantage of hydrogen is that it is a clean and renewable energy source/carrier with high specific heat of combustion and no contribution to the Greenhouse effect, and can be used in many industrial applications. This review discusses fermentative hydrogen production from various points of view. First, the theoretical principles of the biological processes taking place in hydrogen production, as well as the organisms responsible for this process, are described. Second, practical aspects of fermentative hydrogen production are overviewed. Suitable conditions for the hydrogen-producers (pH and temperature), suitable substrates for hydrogen production and applicable reactor designs are discussed. Finally, the challenges faced by fermentative hydrogen production are discussed. Current research directions are listed together with the most important problems currently constraining full-scale applicatio

Supplementation of cobalt to UASB reactors by pulse dosing: CoCl2 versus CoEDTA(2-) pulses

The effect of chelation on the dosing strategy of cobalt to restore the performance of a cobalt limited methanol-fed bioreactor was investigated. Three upflow anaerobic sludge bed (UASB) reactors (30 degrees C, pH 7.0) were operated with methanol as the substrate at an organic loading rate of 8.5 g COD L-1 d(-1). One UASB reactor was supplied with several pulses of cobalt bound to EDTA, and its operation was compared to that of another UASB reactor to which several pulses Of CoCl2 were given. The addition of cobalt (5 mu moles cobalt per litre of reactor volume) in the form of CoCl2 creates a pool of cobalt in the granular sludge matrix due to the high cobalt retention (around 90%). The methanogens present in the granular sludge are able to use that cobalt pool for stable methane formation from methanol during the subsequent 15 days. When added as Co-EDTA(2-), only around 8% of the cobalt added is retained. The small amount of retained cobalt in case of Co-EDTA(2-) addition supports methylotrophic methanogenesis only a few operational days. Furthermore, the side-effects EDTA has on the granule matrix or microbial cells make EDTA an unsuitable ligand for cobalt dosage in full-scale applications

Zinc deprivation of methanol fed anaerobic granular sludge bioreactors

The effect of omitting zinc from the influent of mesophilic (30 degrees C) methanol fed upflow anaerobic sludge bed (UASB) reactors, and latter zinc supplementation to the influent to counteract the deprivation, was investigated by coupling the UASB reactor performance to the microbial ecology of the bioreactor sludge. Limitation of the specific methanogenic activity (SMA) on methanol due to the absence of zinc from the influent developed after 137 days of operation. At that day, the SMA in medium with a complete trace metal solution except Zn was 3.4 g CH4-COD g VSS-1 day(-1), compared to 4.2 g CH4-COD g VSS-1 day(-1) in a medium with a complete (including zinc) trace metal solution. The methanol removal capacity during these 137 days was 99% and no volatile fatty acids accumulated. Two UASB reactors, inoculated with the zinc-deprived sludge, were operated to study restoration of the zinc limitation by zinc supplementation to the bioreactor influent. In a first reactor, no changes to the operational conditions were made. This resulted in methanol accumulation in the reactor effluent after 12 days of operation, which subsequently induced acetogenic activity 5 days after the methanol accumulation started. Methano-genesis could not be recovered by the continuous addition of 0.5 mu M ZnCl2 to the reactor for 13 days. In the second reactor, 0.5 mu M ZnCl2 was added from its start-up. Although the reactor stayed 10 days longer methanogenically than the reactor operated without zinc, methanol accumulation was observed in this reactor (up to 1.1 g COD-MeOH L-1) as well. This study shows that zinc limitation can induce failure of methanol fed UASB reactors due to acidification, which cannot be restored by resuming the continuous supply of the deprived metal

Divalent metal addition restores sulfide-inhibited N2O reduction in Pseudomonas aeruginosa

Hydrogen sulfide (H2S) inhibits the last step of the denitrification process, i.e. the reduction of nitrous oxide (N2O) to dinitrogen gas (N-2), both in natural environments (marine sediments) and industrial processes (activated sludge, methanogenic sludge, BioDeNOx process). In a previously published study, we showed that the inhibitory effect of sulfide to N2O reduction in mixed microbial communities is reversible and can be counteracted by dosing trace amounts of copper. It remained, however, unclear if this was due to copper sulfide precipitation or a retrofitting of the copper containing N2O-reductase (N2OR). The present study aimed to elucidate the mechanism of the restoration of sulfide-inhibited N2O reducing activity by metal addition to a pure Pseudomonas aeruginosa culture. This was done by using other metals (zinc, cobalt and iron) in comparison with copper. Zinc and cobalt clearly alleviated the sulfide inhibition of N2OR to the same extent as copper and the activity restoration was extremely fast (within 15 min, Fig. 3) for zinc, cobalt and copper. This suggests that the alleviation of the inhibitory effect of sulfide is due to metal sulfide precipitation and thus not exclusively limited to Cu. This work also underlines the importance of metal speciation: supply of iron did not restore the N2OR activity because it was precipitated by the phosphates present in the medium and thus could not precipitate the sulfide