The effect of incubation temperature on steady-state concentrations of hydrogen and volatile fatty acids during anaerobic degradation in slurries from wetland sediments

Abstract Increasing the incubation temperature of two swamp slurries from 2°C to37°C resulted in a 8- to 18-fold increase in the H₂ partial pressure. The concentration of volatile fatty acids remained fairly constant except for butyrate, which decreased with increasing temperature. Calculation of Gibbs free energies of syntrophic degradation of butyrate and propionate, and of methanogenesis from acetate and H₂ revealed that these reactions were exergonic after the slurries had stabilized at the incubation temperatures. The changes in H₂ partial pressure and butyrate concentration with temperature were found important to render the processes exergonic within the tested temperature range.     

Biogenic methane production in formation waters from a large gas field in the North Sea

Methanogenesis was investigated in formation waters from a North Sea oil rimmed gas accumulation containing biodegraded oil, which has not been subject to seawater injection. Activity and growth of hydrogenotrophic methanogens was measured but acetoclastic methanogenesis was not detected. Hydrogenotrophic methanogens showed activity between 40 and 80°C with a temperature optimum (ca. 70°C) consistent with in situ reservoir temperatures. They were also active over a broad salinity range, up to and consistent with the high salinity of the waters (90 g l⁻¹). These findings suggest the methanogens are indigenous to the reservoir. The conversion of H₂ and CO₂ to CH₄ in methanogenic enrichments was enhanced by the addition of inorganic nutrients and was correlated with cell growth. Addition of yeast extract also stimulated methanogenesis. Archaeal 16S rRNA gene sequences recovered from enrichment cultures were closely related to Methanothermobacter spp. which have been identified in other high-temperature petroleum reservoirs. It has recently been suggested that methanogenic oil degradation may be a major factor in the development of the world’s heavy oils and represent a significant and ongoing process in conventional deposits. Although an oil-degrading methanogenic consortium was not enriched from these samples the presence and activity of communities of fermentative bacteria and methanogenic archaea was demonstrated. Stimulation of methanogenesis by addition of nutrients suggests that in situ methanogenic biodegradation of oil could be harnessed to enhance recovery of stranded energy assets from such petroleum systems.     

Growth of Syntrophomonas wolfei in pure culture on crotonate

A Synthrophomonas wolfei-Methanospirillum hungatei coculture was adapted to catabolize crotonate. S. wolfei was then isolated in axenic culture using agar spread plates and roll tubes with crotonate as the sole energy source. S. wolfei catabolized crotonate via a disproportionation mechanism similar to that of some Clostridium species. Growth on crotonate was very slow (specific growth rate of 0.029 h^–1) but the conversion of energy into cell material was very efficient with cell yields of 14.6 g (dry wt.) per mol of crotonate. S. wolfei alone did not catabolize butyrate, but butyrate was stoichiometrically degraded to acetate and presumably methane when S. wolfei was reassociated with M. hungatei. S. wolfei-M. hungatei cocultures accumulated some butyrate during growth on crotonate indicating that protons were not the sole electron acceptors used for crotonate oxidation by the coculture.     

Methanofuran-b is required for CO2 formation from formaldehyde by Methanosarcina barkeri

Abstract Extracts of Methanosarcina barkeri strain Fasaro oxidized formaldehyde to CO₂ with methyl-coenzyme M as the natural terminal electron acceptor resulting in methanogenesis. A combination of the artificial electron acceptors methylviologen and metronidazole could substitute for methyl-coenzyme M. The rate of formaldehyde oxidation was thereby increased. Taking advantage of this artificial electron acceptor system the role of cofactors in formaldehyde oxidation was investigated. Cofactor-free extract of M. barkeri did not catalyze the oxidation of formaldehyde. CO₂ formation could be restored by the addition of tetrahydromethanopterin-b (H₄MPT-b) and methanofuran-b (MFR-b) from M. barkeri . Other low molecular weight or heat-resistant compounds stimulating formaldehyde oxidation were not found. Formaldehyde oxidation seems, therefore, to proceed via H₄ MPT-b and MFR-b-derivatives already shown to be involved in methanogenesis from H₂+ CO₂.     

Characterization of an alkane-degrading methanogenic enrichment culture from production water of an oil reservoir after 274 days of incubation

Oil reservoirs represent special habitats for the activity of anaerobic microbial communities in the transformation of organic compounds. To understand the function of microbial communities in oil reservoirs under anaerobic conditions, an alkane-degrading methanogenic enrichment culture was established and analyzed. Results showed that a net 538 ??mol of methane higher than the controls were produced over 274 days of incubation in microcosms amended with alkanes and a decrease in the alkanes profile was also observed. Phylogenetic analysis of 16S rRNA gene sequences retrieved from the enrichment microcosms indicated that the archaeal phylotypes were mostly related to members of the orders Methanobacteriales and Methanosarcinales. The bacterial clone library was composed of sequences affiliated with the Firmicutes, Proteobacteria, Deferribacteres, and Bacteroidetes. However, most of the bacterial clones retrieved from the enrichment cultures showed low similarity to 16S rRNA gene sequences of the cultured members, indicating that the enrichment cultures contained novel bacterial species. Though alkane-degrading methanogenic enrichment consortium has rarely been reported from petroleum reservoirs, our results indicated that oilfield production water harbors a microbial community capable of syntrophic conversion of n-alkanes to methane, which sheds light on the bio-utilization of marginal oil reservoirs for enhanced energy recovery.     

Organics, sulfates and ammonia removal from acrylic fiber manufacturing wastewater using a combined Fenton-UASB (2 phase)-SBR system

A combined Fenton-UASB (2 phase)-SBR system was employed to treat acrylic fiber manufacturing wastewater. The Chemical Oxygen Demand (COD) removal and effluent Biochemical Oxygen Demand (BOD) to COD were 65.5% and 0.529%, respectively, with the optimal Fenton conditions: ferrous was 300 mg/L; hydrogen peroxide was 500 mg/L; pH was 3.0; reaction time was 2.0 h. In two-phase UASB reactor, mesophilic operation (35±0.5 °C) was performed with hydraulic retention time (HRT) varied between 28 and 40 h. The results showed that with the HRT not less than 38 h, COD and sulfate removal were 65% and 75%, respectively. The greatest sizes of granule formed in the sulfate-reducing and methane-producing phases were 5 and 2 mm, respectively. Sulfate-reducing bacteria (SRB) accounted for 35% in the sulfate-reducing phase while methane-producing archaea (MPA) accounted for 72% in the methane-producing phase. During the SBR process, shortcut nitrification was achieved by temperature control of 30 °C.     

Effect of low solids retention time and focused pulsed pre-treatment on anaerobic digestion of waste activated sludge

The interacting effects of Focused Pulsed (FP) treatment and solids retention time (SRT) were evaluated in laboratory-scale digesters operated at SRTs of 2-20 days. Anaerobic digestion and methanogenesis of waste activated sludge (WAS) were stable for SRT ? 5 days, but the effluent soluble organic compounds increased significantly for SRT = 2 days due to a combination of faster hydrolysis kinetics and washout of methanogens. FP treatment increased the CH₄ production rate and TCOD removal efficiency by up to 33% and 18%, respectively, at a SRT of 20 days. These effects were the result of an increase in the hydrolysis rate, since the concentrations of soluble components remained low for SRT ? 5 days. Alternately, FP pre-treatment of WAS allowed the same conversion of TCOD to CH₄ with a smaller SRT and digester size: e.g., 40% size savings with a CH₄ conversion of 0.23 g CH₄-COD/g COD_in.     

Methanogenic pathways in Methanosphaera stadtmanae

Methanosphaera stadtmanae reduces methanol to CH4 in a similar way as Methanosarcina barkeri. Low activities of 5,10-methylenetetrahydromethanopterin dehydrogenase (MTDH) and reductase (MTR) were found. From studies on formaldehyde oxidation and reduction it was concluded that most likely the inability to reduce CO2 to CH4 was due to the lack of an active or the presence of an inactive CO2 reductase system and methyltetrahydromethanopterin (methyl-H4MPT): coenzyme M methyltransferase. Methanofuran was not detected, while the presence of a pterin, analogous to H4MPT, could be substantiated from its degradation products in boiled extracts.