Using enriched cultures for elevation of anaerobic syntrophic interactions between acetogens and methanogens in a high-load continuous digester

Volatile fatty acids (VFAs) are key intermediates in anaerobic digestion. Enriched acetogenic and methanogenic cultures were used for syntrophic anaerobic digestion of VFAs in a high-load continuous reactor fed with acetic (HAc), propionic (HPr) and butyric (HBu) acids at maximum concentrations of 5, 3 and 4 g/L, respectively. Interactive effects of HPr, HBu and HAc were analyzed. Furthermore, hydraulic retention time (HRT) and methanogen to acetogen population ratio (M/A) were investigated as key microbiological and operating variables of VFA anaerobic degradations. Optimum conditions were found to be HPr = 1125.0 mg/L, HBu = 1833.4 mg/L, HAc = 1727.4 mg/L, HRT = 21 h and M/A = 2.5 (corresponding to maximum VFA removal and biogas production rate (BPR)). Results of verification experiments and predicted values from fitted correlations were in close agreement at 95% confidence interval. HRT and M/A had positive effects on VFA removal and BPR. M/A was the most important factor that affected BPR. All VFAs inhibited VFA removals.     

Copper stressed anaerobic fermentation: biogas properties,process stability,biodegradation and enzyme responses

The effect of copper (added as CuCl₂) on the anaerobic co-digestion of Phragmites straw and cow dung was studied in pilot experiments by investigating the biogas properties, process stability, substrate degradation and enzyme activities at different stages of mesophilic fermentation. The results showed that 30 and 100 mg/L Cu²⁺ addition increased the cumulative biogas yields by up to 43.62 and 20.77% respectively, and brought forward the daily biogas yield peak, while 500 mg/L Cu²⁺ addition inhibited biogas production. Meanwhile, the CH₄ content in the 30 and 100 mg/L Cu²⁺-added groups was higher than that in the control group. Higher pH values (close to pH 7) and lower oxidation–reduction potential (ORP) values in the Cu²⁺-added groups after the 8th day indicated better process stability compared to the control group. In the presence of Cu²⁺, the degradation of volatile fatty acids (VFAs) and other organic molecules (represented by chemical oxygen demand, COD) generated from hydrolysis was enhanced, and the ammonia nitrogen (NH₄ ⁺-N) concentrations were more stable than in the control group. The contents of lignin and hemicellulose in the substrate declined in the Cu²⁺-added groups while the cellulose contents did not. Neither the cellulase nor the coenzyme F₄₂₀ activities could determine the biogas producing efficiency. Taking the whole fermentation process into account, the promoting effect of Cu²⁺ addition on biogas yields was mainly attributable to better process stability, the enhanced degradation of lignin and hemicellulose, the transformation of intermediates into VFA, and the generation of CH₄ from VFA.     

Comparative biochemical analysis during the anaerobic digestion of lignocellulosic biomass from six morphological parts of Williams Cavendish banana (Triploid Musa AAA group) plants

We studied banana lignocellulosic biomass (BALICEBIOM) that is abandoned after fruit harvesting, and assessed its biochemical methane potential, because of its potential as an energy source. We monitored biogas production from six morphological parts (MPs) of the “Williams Cavendish” banana cultivar using a modified operating procedure (KOP) using KOH. Volatile fatty acid (VFA) production was measured using high performance liquid chromatography. The bulbs, leaf sheaths, petioles–midribs, leaf blades, rachis stems, and floral stalks gave total biogas production of 256, 205, 198, 126, 253, and 221 ml g^?1 dry matter, respectively, and total biomethane production of 150, 141, 127, 98, 162, and 144 ml g^?1, respectively. The biogas production rates and yields depended on the biochemical composition of the BALICEBIOM and the ability of anaerobic microbes to access fermentable substrates. There were no significant differences between the biogas analysis results produced using KOP and gas chromatography. Acetate was the major VFA in all the MP sample culture media. The bioconversion yields for each MP were below 50 %, showing that these substrates were not fully biodegraded after 188 days. The estimated electricity that could be produced from biogas combustion after fermenting all of the BALICEBIOM produced annually by the Cameroon Development Corporation–Del Monte plantations for 188 days is approximately 10.5 × 10⁶ kW h (which would be worth 0.80–1.58 million euros in the current market). This bioenergy could serve the requirements of about 42,000 people in the region, although CH₄ productivity could be improved.     

Using Contaminated Plants Involved in Phytoremediation for Anaerobic Digestion

This study investigated the anaerobic digestion capability of five plants and the effects of copper (Cu) and S,S’-ethylenediaminedisuccinic acid (EDDS, a chelator widely used in chelant-assisted phytoremediation) on biogas production to determine a feasible disposal method for plants used in remediation. The results showed that in addition to Phytolacca americana L., plants such as Zea mays L., Brassica napus L., Elsholtzia splendens Nakai ex F. Maekawa, and Oenothera biennis L. performed well in biogas production. Among these, O. biennis required the shortest period to finish anaerobic digestion. Compared to normal plants with low Cu content, the plants used in remediation with increased Cu levels (100 mg kg^?1) not only promoted anaerobic digestion and required a shorter anaerobic digestion time, but also increased the methane content in biogas. When the Cu content in plants increased to 500, 1000, and 5000 mg kg^?1, the cumulative biogas production decreased by 12.3%, 14.6%, and 41.2%, respectively. Studies also found that EDDS conspicuously restrained biogas production from anaerobic digestion. The results suggest that anaerobic digestion has great potential for the disposal of contaminated plants and may provide a solution for the resource utilization of plants used in remediation.     

Volatile fatty acids productivity by anaerobic co-digesting waste activated sludge and corn straw: Effect of feedstock proportion

Volatile fatty acids (VFAs) are the most suitable and biodegradable carbon substrates for many bioprocesses. This study explored a new approach to improve the VFAs production from anaerobic co-digesting waste activated sludge (WAS) with corn straw (CS). The effect of feedstock proportion on the acidification efficiency was investigated. The maximum VFAs yield (corresponding fermentation time) was substantially increased 69% (96 h), 45% (72 h), 13% (120 h) and 12% (120 h) with 50%, 35%, 25% and 20% CS proportion of feedstock, respectively. HAc (acetic acid) was consistently the most abundant, followed by HPr (propionic acid) and n-HBu (butyric acid) in the co-digesting tests. The increase of CS in feedstock led to more production of HAc and HPr. Moreover, the consumption of protein and carbohydrate were also improved remarkably from 2955 and 249 mg COD/L (individual WAS fermentation) to 6575 and 815 mg COD/L (50%_WAS:50%_CS co-digestion) from 120 onward, respectively. The highest contribution of CS to additional VFAs production was1113 mg VFAs (as COD)/g CS/L in the 65%_WAS:35%_CS co-digesting test. Our study indicated a valuable method to improve VFAs production from anaerobic co-digesting WAS and CS.     

Volatile fatty acids (VFAs) and methane from food waste and cow slurry: Comparison of biogas and VFA fermentation processes

The potential of various biomasses for the production of green chemicals is currently one of the key topics in the field of the circular economy. Volatile fatty acids (VFAs) are intermediates in the methane formation pathway of anaerobic digestion and they can be produced in similar reactors as biogas to increase the productivity of a digestion plant, as VFAs have more varying end uses compared to biogas and methane. In this study, the aim was to assess the biogas and VFA production of food waste (FW) and cow slurry (CS) using the anaerobic biogas plant inoculum treating the corresponding substrates. The biogas and VFA production of both biomasses were studied in identical batch scale laboratory conditions while the process performance was assessed with chemical and microbial analyses. As a result, FW and CS were shown to have different chemical performances and microbial dynamics in both VFA and biogas processes. FW as a substrate showed higher yields in both processes (435 ml CH₄/g VS_fed and 434 mg VFA/g VS_fed) due to its characteristics (pH, organic composition, microbial communities), and thus, the vast volume of CS makes it also a relevant substrate for VFA and biogas production. In this study, VFA profiles were highly dependent on the substrate and inoculum characteristics, while orders Clostridiales and Lactobacillales were connected with high VFA and butyric acid production with FW as a substrate. In conclusion, anaerobic digestion supports the implementation of the waste management hierarchy as it enables the production of renewable green chemicals from both urban and rural waste materials.     

Interactions between substrate,fermentation end-products,buffering systems and gas production upon fermentation of different carbohydrates by mixed rumen microorganisms in vitro

Summary In animal nutrition, incubation of feed samples with CO₂/HCO₃-buffered rumen fluid is used to predict the nutritional values of the feed. During fermentation, volatile fatty acids (VFAs) are produced, which release CO₂ from the buffer through their H⁺ ions. This indirect gas production amounted to 20.8 ml gas per mmol VFA. By incubating glucose, rice starch and cellulose, the relationship between direct and indirect gas production in relation to fermentation kinetics was studied. The total amount of gas formed was found to be dependent on the composition of the fermentation end-products formed. This could be described by: ml gas = M_v·mmol HAc + 2M_v·mmol HB + 0.87M_v·mmol Tot. VFA where HAc = acetic acid; HB = butyric acid; and M_v = molar gas volume. No clear relationship was found between the rate of fermentation and total gas production. From rice starch more total gas was produced than from glucose and cellulose, which were fermented faster and slower, respectively. Correspondence to: S. F. Spoelstra     

Electrolytic extraction drives volatile fatty acid chain elongation through lactic acid and replaces chemical pH control in thin stillage fermentation

Background

Volatile fatty acids (VFA) are building blocks for the chemical industry. Sustainable, biological production is constrained by production and recovery costs, including the need for intensive pH correction. Membrane electrolysis has been developed as an in situ extraction technology tailored to the direct recovery of VFA from fermentation while stabilizing acidogenesis without caustic addition. A current applied across an anion exchange membrane reduces the fermentation broth (catholyte, water reduction: H₂O + e^? → ½ H₂ + OH^?) and drives carboxylate ions into a clean, concentrated VFA stream (anolyte, water oxidation: H₂O → 2e^? + 2 H⁺ + O₂).

Results

In this study, we fermented thin stillage to generate a mixed VFA extract without chemical pH control. Membrane electrolysis (0.1 A, 3.22 ± 0.60 V) extracted 28 ± 6 % of carboxylates generated per day (on a carbon basis) and completely replaced caustic control of pH, with no impact on the total carboxylate production amount or rate. Hydrogen generated from the applied current shifted the fermentation outcome from predominantly C2 and C3 VFA (64 ± 3 % of the total VFA present in the control) to majority of C4 to C6 (70 ± 12 % in the experiment), with identical proportions in the VFA acid extract. A strain related to Megasphaera elsdenii (maximum abundance of 57 %), a bacteria capable of producing mid-chain VFA at a high rate, was enriched by the applied current, alongside a stable community of Lactobacillus spp. (10 %), enabling chain elongation of VFA through lactic acid. A conversion of 30 ± 5 % VFA produced per sCOD fed (60 ± 10 % of the reactive fraction) was achieved, with a 50 ± 6 % reduction in suspended solids likely by electro-coagulation.

Conclusions

VFA can be extracted directly from a fermentation broth by membrane electrolysis. The electrolytic water reduction products are utilized in the fermentation: OH^? is used for pH control without added chemicals, and H₂ is metabolized by species such as Megasphaera elsdenii to produce greater value, more reduced VFA. Electro-fermentation displays promise for generating added value chemical co-products from biorefinery sidestreams and wastes.

     

Co-digestion of manure and whey for in situ biogas upgrading by the addition of H2: process performance and microbial insights

In situ biogas upgrading was conducted by introducing H₂ directly to the anaerobic reactor. As H₂ addition is associated with consumption of the CO₂ in the biogas reactor, pH increased to higher than 8.0 when manure alone was used as substrate. By co-digestion of manure with acidic whey, the pH in the anaerobic reactor with the addition of hydrogen could be maintained below 8.0, which did not have inhibition to the anaerobic process. The H₂ distribution systems (diffusers with different pore sizes) and liquid mixing intensities were demonstrated to affect the gas-liquid mass transfer of H₂ and the biogas composition. The best biogas composition (75:6.6:18.4) was obtained at stirring speed 150 rpm and using ceramic diffuser, while the biogas in the control reactor consisted of CH₄ and CO₂ at a ratio of 55:45. The consumed hydrogen was almost completely converted to CH₄, and there was no significant accumulation of VFA in the effluent. The study showed that addition of hydrogen had positive effect on the methanogenesis, but had no obvious effect on the acetogenesis. Both hydrogenotrophic methanogenic activity and the concentration of coenzyme F₄₂₀ involved in methanogenesis were increased. The archaeal community was also altered with the addition of hydrogen, and a Methanothermobacter thermautotrophicus related band appeared in a denaturing gradient gel electrophoresis gel from the sample of the reactor with hydrogen addition. Though the addition of hydrogen increased the dissolved hydrogen concentration, the degradation of propionate was still thermodynamically feasible at the reactor conditions.     

Medium chain length polyhydroxyalkanoate (mcl-PHA) production from volatile fatty acids derived from the anaerobic digestion of grass

A two step biological process for the conversion of grass biomass to the biodegradable polymer medium chain length polyhydroxyalkanoate (mcl-PHA) was achieved through the use of anaerobic and aerobic microbial processes. Anaerobic digestion (mixed culture) of ensiled grass was achieved with a recirculated leach bed bioreactor resulting in the production of a leachate, containing 15.3 g/l of volatile fatty acids (VFAs) ranging from acetic to valeric acid with butyric acid predominating (12.8 g/l). The VFA mixture was concentrated to 732.5 g/l with a 93.3 % yield of butyric acid (643.9 g/l). Three individual Pseudomonas putida strains, KT2440, CA-3 and GO16 (single pure cultures), differed in their ability to grow and accumulate PHA from VFAs. P. putida CA-3 achieved the highest biomass and PHA on average with individual fatty acids, exhibited the greatest tolerance to higher concentrations of butyric acid (up to 40 mM) compared to the other strains and exhibited a maximum growth rate (μ_MAX?=?0.45 h^?1). Based on these observations P. putida CA-3 was chosen as the test strain with the concentrated VFA mixture derived from the AD leachate. P. putida CA-3 achieved 1.56 g of biomass/l and accumulated 39 % of the cell dry weight as PHA (nitrogen limitation) in shake flasks. The PHA was composed predominantly of 3-hydroxydecanoic acid (>65 mol%).     

Intensifying Clean Energy Production Through Cultivating Mixotrophic Microalgae from Digestates of Biogas Systems: Effects of Light Intensity,Medium Dilution,and Cultivating Time

High-strength wastewaters after being digested for biogas production in anaerobic digesters still contain substantial nutrients and organics. The anaerobic digestates from four major industries in Thailand were tested with batch cultivation of Chlorella sp. for oil production potentials. Pig farm digestate was found most suitable as the growth medium generating 0.95 g/L_medium (dry biomass), which was 1.16–3.06 times of other digestates tested. Considerable removals of nitrogen and phosphorus achieved were an added benefit to the goal of ultimate treatment of these wastewaters. Light intensity had strong influence on growth and heterotrophic metabolism up to 78 μmol/m²/s, while the dilution of digestate above 2.4× diminished growth potential and lipid production. A quadratic regression model was constructed to describe interaction of light intensity, dilution factor, and time of cultivation to lipid production with a satisfactory precision. Light intensity could influence fatty acid composition, although palmitic acid was found predominant at 47.1 %. The algae oil generated could potentially increase the total energy output from anaerobic digesters of a typical pig farm by 22 %.     

Biogas properties and enzymatic analysis during anaerobic fermentation of <Emphasis Type="Italic">Phragmites australis</Emphasis> straw and cow dung: influence of nickel chloride supplement

The importance of nickel (added as NiCl₂) on mesophilic anaerobic fermentation of Phragmites australis straw and cow dung was demonstrated by investigating the biogas properties, pH values, organic matter degradation [chemical oxygen demand (COD)] and enzyme activities (cellulase, protease and dehydrogenase) during the fermentation process. The results showed that Ni²⁺ addition increased the cumulative biogas yields by >18 % by improving the efficiency of first peak stage and bringing forward the second peak stage. The pH values were not significantly influenced by Ni²⁺ addition (p > 0.05). Biogas yields were associated with variations in COD concentrations rather than momentary concentrations. At the start-up stage of fermentation (4th day), the biogas yields increased gradually together with the increase of dehydrogenase activities at elevated Ni²⁺ concentrations when cellulase and protease activities were similar in all test groups. It is suggested that Ni²⁺ addition was mainly dependent on the methanogenic stage. After the start-up stage, the impact of Ni²⁺ addition on biogas production was mainly dependent on its effect on cellulase activities, rather than protease or dehydrogenase activities.     

Degradation of oxytetracycline and its impacts on biogas-producing microbial community structure

The effect of veterinary antibiotics in anaerobic digesters is a concern where methane production efficiency is highly dependent on microbial community structure. In this study, both anaerobic degradation of a common veterinary antibiotic, oxytetracycline (OTC), and its effects on an anaerobic digester microbial community were investigated. Qualitative and quantitative molecular tools were used to monitor changes in microbial community structure during a 60-day batch incubation period of cow manure with the addition of different concentrations of the antibiotic. Molecular data were interpreted by a further redundancy analysis as a multivariate statistics approach. At the end of the experiment, approximately 48, 33, and 17 % of the initially added 50, 100, and 200 mg l^?1 of OTC was still present in the serum bottles which reduced the biogas production via accumulation of some of the volatile fatty acids (VFAs). Biogas production was highly correlated with Methanobacteriales and Methanosarcinales gene copy numbers, and those parameters were negatively affected with oxytetracycline and VFA concentrations.     

Improving biogas production from protein-rich distillery wastewater by decreasing ammonia inhibition

A large quantity of protein-rich distillery wastewater is produced during the process of bio-ethanol production from kitchen waste. It is difficult, however, to treat protein-rich distillery wastewater by anaerobic digestion due to ammonia inhibition. In this study, a novel method was investigated to reduce ammonia inhibition during thermophilic anaerobic digestion through the recirculation of water-washed biogas into the headspace (R1 system) or liquid phase (R2 system) of the reactors. The results show that the method greatly improved biogas production from distillery wastewater. R2 system achieved stable biogas production at a higher organic loading rate (OLR) of 4.0 g VTS/L/d than R1 system at 3.0 g VTS/L/d. At the same OLR, we observed a higher biogas production rate but lower accumulation of NH₄⁺ and volatile fatty acids in the reactor, and higher ammonia absorption rate in the water tank of R2 system than R1 system. The better performance of R2 system could be attributed to the more efficient removal of ammonia from liquid phase. In addition, adjusting the C/N ratio of distillery wastewater from 9.0 to 11.4 significantly enhanced the maximum OLR from 3.0 to 7.0 g VTS/L/d in R1 system.     

Composition and dynamics of microbial community in a zeolite biofilter-membrane bioreactor treating coking wastewater

In this study, a lab-scale anaerobic/anoxic/zeolite biofilter-membrane bioreactor (A₁/A₂/ZB-MBR) was designed to treat coking wastewater. The 454 pyrosequencing was used to obtain the composition and dynamics of microbial community about the treatment system. The results showed that the system yielded stable effluent chemical oxidation demand (158.5?±?21.8 mg/L) and ammonia (8.56?±?7.30 mg/L), but fluctuant total nitrogen (31.4–165.1 mg/L) concentrations. In addition, 66,256 16S rRNA gene sequences were obtained from A₂ and ZB-MBR, and the microbial diversity and richness for five samples were determined. Although community compositions in the five samples were quite different, bacteria assigned to phylum Proteobacteria and class Flavobacteria commonly existed and dominated the microbial populations. The pyrosequencing analysis revealed that the microbial community shifted in the ZB-MBR with the presence of zeolite. Some taxa began to appear in ZB-MBR and contributed to the system performance. Additionally, Nitrosomonas and Nitrobacter gradually became the dominant ammonia-oxidizing bacteria and nitrite-oxidizing bacteria during the operation, respectively, which are favorable for the stabilized ammonia removal. Our results proved that the ZB-MBR is an alternative technique for treating coking wastewater.     

The effect of temperature and retention time on methane production and microbial community composition in staged anaerobic digesters fed with food waste

Background

Food waste is a large bio-resource that may be converted to biogas that can be used for heat and power production, or as transport fuel. We studied the anaerobic digestion of food waste in a staged digestion system consisting of separate acidogenic and methanogenic reactor vessels. Two anaerobic digestion parameters were investigated. First, we tested the effect of 55 vs. 65 °C acidogenic reactor temperature, and second, we examined the effect of reducing the hydraulic retention time (HRT) from 17 to 10 days in the methanogenic reactor. Process parameters including biogas production were monitored, and the microbial community composition was characterized by 16S amplicon sequencing.

Results

Neither organic matter removal nor methane production were significantly different for the 55 and 65 °C systems, despite the higher acetate and butyrate concentrations observed in the 65 °C acidogenic reactor. Ammonium levels in the methanogenic reactors were about 950 mg/L NH₄ ⁺ when HRT was 17 days but were reduced to 550 mg/L NH₄ ⁺ at 10 days HRT. Methane production increased from ~ 3600 mL/day to ~ 7800 when the HRT was decreased. Each reactor had unique environmental parameters and a correspondingly unique microbial community. In fact, the distinct values in each reactor for just two parameters, pH and ammonium concentration, recapitulate the separation seen in microbial community composition. The thermophilic and mesophilic digesters were particularly distinct from one another. The 55 °C acidogenic reactor was mainly dominated by Thermoanaerobacterium and Ruminococcus, whereas the 65 °C acidogenic reactor was initially dominated by Thermoanaerobacterium but later was overtaken by Coprothermobacter. The acidogenic reactors were lower in diversity (34–101 observed OTU_0.97, 1.3–2.5 Shannon) compared to the methanogenic reactors (472–513 observed OTU_0.97, 5.1–5.6 Shannon). The microbial communities in the acidogenic reactors were > 90% Firmicutes, and the Euryarchaeota were higher in relative abundance in the methanogenic reactors.

Conclusions

The digestion systems had similar biogas production and COD removal rates, and hence differences in temperature, NH₄ ⁺ concentration, and pH in the reactors resulted in distinct but similarly functioning microbial communities over this range of operating parameters. Consequently, one could reduce operational costs by lowering both the hydrolysis temperature from 65 to 55 °C and the HRT from 17 to 10 days.

     

Improved biogas production from palm oil mill effluent by a scaled-down anaerobic treatment process

This is a scale-down study of a 500-m³ methane recovery test plant for anaerobic treatment of palm oil mill effluent (POME) where biomass washout has become one of the problems because of the continuous mixing of effluent during anaerobic treatment of POME. Therefore, in this study, anaerobic POME treatment using a scaled down 50-l bioreactor which mimicked the 500-m³ bioreactor was carried out to improve biogas production with and without biomass sedimentation. Three sets of experiments were conducted under different conditions in terms of biomass sedimentation applied to the system. The first experiment was operated under semi-continuous mode whereas the second and third experiments were operated based on mix and settle mode. As expected, biomass retention improved the anaerobic process as the POME treatment incorporated with mix and settle system were able to operate at an organic loading rate (OLR) of 3.5 and 6.0 kg COD/m³/day respectively, while the semi-continuous operated anaerobic treatment only achieved OLR of 3.0 kg COD/m³/day. The highest biogas and methane production rates achieved were 2.42 m³/m³ of reactor/day and 0.992 m³/m³ of reactor/day, respectively at OLR 6.0 kg COD/m³/day. The biomass or solids retention in the reactors was represented by the total solids measured in this study.     

Integrated biogas upgrading and hydrogen utilization in an anaerobic reactor containing enriched hydrogenotrophic methanogenic culture

Biogas produced by anaerobic digestion, is mainly used in a gas motor for heat and electricity production. However, after removal of CO₂, biogas can be upgraded to natural gas quality, giving more utilization possibilities, such as utilization as autogas, or distant utilization by using the existing natural gas grid. The current study presents a new biological method for biogas upgrading in a separate biogas reactor, containing enriched hydrogenotrophic methanogens and fed with biogas and hydrogen. Both mesophilic‐ and thermophilic anaerobic cultures were enriched to convert CO₂ to CH₄ by addition of H₂. Enrichment at thermophilic temperature (55°C) resulted in CO₂ and H₂ bioconversion rate of 320 mL CH₄/(gVSS h), which was more than 60% higher than that under mesophilic temperature (37°C). Different dominant species were found at mesophilic‐ and thermophilic‐enriched cultures, as revealed by PCR–DGGE. Nonetheless, they all belonged to the order Methanobacteriales, which can mediate hydrogenotrophic methanogenesis. Biogas upgrading was then tested in a thermophilic anaerobic reactor under various operation conditions. By continuous addition of hydrogen in the biogas reactor, high degree of biogas upgrading was achieved. The produced biogas had a CH₄ content, around 95% at steady‐state, at gas (mixture of biogas and hydrogen) injection rate of 6 L/(L day). The increase of gas injection rate to 12 L/(L day) resulted in the decrease of CH₄ content to around 90%. Further study showed that by decreasing the gas–liquid mass transfer by increasing the stirring speed of the mixture the CH₄ content was increased to around 95%. Finally, the CH₄ content around 90% was achieved in this study with the gas injection rate as high as 24 L/(L day). Biotechnol. Bioeng. 2012; 109: 2729–2736. © 2012 Wiley Periodicals, Inc.     

Bioaugmentation of Syntrophic Acetate-Oxidizing Culture in Biogas Reactors Exposed to Increasing Levels of Ammonia

The importance of syntrophic acetate oxidation for process stability in methanogenic systems operating at high ammonia concentrations has previously been emphasized. In this study we investigated bioaugmentation of syntrophic acetate-oxidizing (SAO) cultures as a possible method for decreasing the adaptation period of biogas reactors operating at gradually increased ammonia concentrations (1.5 to 11 g NH₄⁺-N/liter). Whole stillage and cattle manure were codigested semicontinuously for about 460 days in four mesophilic anaerobic laboratory-scale reactors, and a fixed volume of SAO culture was added daily to two of the reactors. Reactor performance was evaluated in terms of biogas productivity, methane content, pH, alkalinity, and volatile fatty acid (VFA) content. The decomposition pathway of acetate was analyzed by isotopic tracer experiments, and population dynamics were monitored by quantitative PCR analyses. A shift in dominance from aceticlastic methanogenesis to SAO occurred simultaneously in all reactors, indicating no influence by bioaugmentation on the prevailing pathway. Higher abundances of Clostridium ultunense and Tepidanaerobacter acetatoxydans were associated with bioaugmentation, but no influence on Syntrophaceticus schinkii or the methanogenic population was distinguished. Overloading or accumulation of VFA did not cause notable dynamic effects on the population. Instead, the ammonia concentration had a substantial impact on the abundance level of the microorganisms surveyed. The addition of SAO culture did not affect process performance or stability against ammonia inhibition, and all four reactors deteriorated at high ammonia concentrations. Consequently, these findings further demonstrate the strong influence of ammonia on the methane-producing consortia and on the representative methanization pathway in mesophilic biogas reactors.     

Anaerobic submerged membrane bioreactor (AnSMBR) treating low-strength wastewater under psychrophilic temperature conditions

An anaerobic submerged membrane bioreactor (AnSMBR) treating low-strength wastewater was operated for 90 days under psychrophilic temperature conditions (20 °C). Besides biogas sparging, additional shear was created by circulating sludge to control membrane fouling. The critical flux concept was used to evaluate the effectiveness of this configuration. Biogas sparging with a gas velocity (U_G) of 62 m/h together with sludge circulation (94 m/h) led to a critical flux of 7 L/(m² h). Nevertheless, a further increase in the U_G only minimally enhanced the critical flux. A low fouling rate was observed under critical flux conditions. The cake layer represented the main fouling resistance after 85 days of operation. Distinctly different volatile fatty acid (VFA) concentrations in the reactor and in the permeate were always observed. This fact suggests that a biologically active part of the cake layer contributes to degrade a part of the daily organic load. Hence, chemical oxygen demand (COD) removal efficiencies of up to 94% were observed. Nevertheless, the biogas balance indicates that even considering the dissolved methane, the methane yield were always lower than the theoretical value, which indicates that the organic compounds were not completely degraded but physically retained by the membrane in the reactor.