Two-stage biogas production by co-digesting molasses wastewater and sewage sludge

We evaluated the feasibility of co-digesting molasses wastewater and sewage sludge in a two-stage hydrogen- and methane-producing system. The highest energy was recovered at the 21-h hydraulic retention time (HRT) of the first hydrogenic reactor and at 56-h HRT of the secondary methanogenic reactor. Hence, the two-stage system recovered 1,822 kJ from 1 L of the mixed wastes (19.7: hydrogenic reactor plus, 1,802 kJ L^?1: methanogenic reactor). Despite the overloaded VFA-run with a short HRT of 56 h, the GAC-CH₄ reactor increased methane production rate and yields due to enhanced pH buffer capacity. An RNA-based community analysis showed that the Ethanoligenens and Methanosaeta dominated the hydrogen and methane bioreactor, respectively. The two-stage system of co-digesting molasses and sewage sludge is particularly cost-effective due to non-pretreatment of sewage sludge.     

Microbial Community Dynamics of a Continuous Mesophilic Anaerobic Biogas Digester Fed with Sugar Beet Silage

The aim of the study was to investigate the long‐term fermentation of an extremely sour substrate without any addition of manure. In the future, the limitation of manure and therefore the anaerobic digestion of silage with a very low buffering capacity will be an increasing general bottleneck for energy production from renewable biomass. During the mesophilic anaerobic digestion of sugar beet silage (without top and leaves) as the sole substrate (without any addition of manure), which had an extreme low pH of around 3.3, the highest specific gas production rate (spec. GPR) of 0.72 L/g volatile solids (VS) d was achieved at a hydraulic retention time (HRT) of 25 days compared to an organic loading rate (OLR) of 3.97 g VS/L d at a pH of around 6.80. The methane (CH₄) content of the digester ranged between 58 and 67 %, with an average of 63 %. The use of a new charge of substrate (a new harvest of the same substrate) with higher phosphate content improved the performance of the biogas digester significantly. The change of the substrate charge also seemed to affect the methanogenic population dynamics positively, thus improving the reactor performance. Using a new substrate charge, a further decrease in the HRT from 25 to 15 days did not influence the digester performance and did not seem to affect the structure of the methanogenic population significantly. However, a decrease in the HRT affected the size of the methanogenic population adversely. The lower spec. GPR of 0.54 L/g VS d attained on day 15 of the HRT could be attributed to a lower size of methanogenic population present in the anaerobic digester during this stage of the process. Furthermore, since sugar beet silage is a relatively poor substrate, in terms of the buffering capacity and the availability of nutrients, an external supply of buffering agents and nutrients is a prerequisite for a safe and stable digester operation.     

Population dynamics of methanogens and methane formation associated with different loading rates of organic acids along with ammonia: redundancy analysis

In anaerobic processes, the population dynamics of methanogens in the methanogenic stage were monitored along with hydraulic retention times (HRTs) shift. Decreasing HRTs increased the loading rates of volatile fatty acids (VFAs) and ammonia. Methanomicrobiales (MMB) began to be dominant at longer than 12.5 days HRT, Methanosarcinales (MSL) were dominant at 8, 10, and 12.5 days HRT, and Methanobacteriales (MBT) were dominant at shorter than 6 days HRT. Increased loading rates of VFAs and ammonia increased MBT, decreased MMB, and had no significant effect on MSL. Maximal daily methane production was observed at 1.57 L/L when MSL copy numbers also reached 3.60 × 10⁷ copy/mL as a peak, which were expressed as positive correlation between DMA and MSL. No sooner had methane yield (MY) increased from 1.15 to 1.32 L/g VS_removed along with HRT reduction from 25 to 22.5 days, then MY gradually decreased from 1.32 to 0.04 L/g VS_removed.     

Some studies on UASB bioreactors for the stabilization of low strength industrial effluents

The suitability of two stage biomethanation process using upflow anaerobic sludge blanket (UASB) bioreactors was studied for the treatment of low strength industrial effluents like rice mill wastewater. Maximum VFA yield was 0.75 mg (as acetic acid) per mg of COD consumed at a flow rate of 25 ml/min. Hydraulic retention time (HRT) of 1 hr was found suitable for acidification process. In the methanogenic reactor, the overall BOD and COD reductions were 89% and 78% respectively at loading rate of 3 kg COD m^х d^у, and HRT of 30 hrs. Gas yield in methanogenic reactor was 0.56 lits. per kg COD consumed which contains 62% v/v methane.     

Enhancement of bioenergy production from organic wastes by two-stage anaerobic hydrogen and methane production process

The present study investigated a two-stage anaerobic hydrogen and methane process for increasing bioenergy production from organic wastes. A two-stage process with hydraulic retention time (HRT) 3 d for hydrogen reactor and 12 d for methane reactor, obtained 11% higher energy compared to a single-stage methanogenic process (HRT 15 d) under organic loading rate (OLR) 3 gVS/(L d). The two-stage process was still stable when the OLR was increased to 4.5 gVS/(L d), while the single-stage process failed. The study further revealed that by changing the HRT_hydrogen:HRT_methane ratio of the two-stage process from 3:12 to 1:14, 6.7%, more energy could be obtained. Microbial community analysis indicated that the dominant bacterial species were different in the hydrogen reactors (Thermoanaerobacterium thermosaccharolyticum-like species) and methane reactors (Clostridiumthermocellum-like species). The changes of substrates and HRT did not change the dominant species. The archaeal community structures in methane reactors were similar both in single- and two- stage reactors, with acetoclastic methanogens Methanosarcina acetivorans-like organisms as the dominant species.     

Hydrogen and methane production through two-stage mesophilic anaerobic digestion of olive pulp

The present study focused on the anaerobic biohydrogen production from olive pulp (two phase olive mill wastes, TPOMW) and the subsequent anaerobic treatment of the effluent for methane production under mesophilic conditions in a two-stage process. Biohydrogen production from water-diluted (1:4) olive pulp was investigated at hydraulic retention times (HRT) of 30 h, 14.5 h and 7.5 h while methane production from the effluent of hydrogenogenic reactor was studied at 20 d, 15 d, 10 d and 5 d HRT. In comparison with previous studies, it has been shown that the thermophilic hydrogen production process was more efficient than the mesophilic one in both hydrogen production rate and yield. The methanogenic reactor was successfully operated at 20, 15 and 10 days HRT while it failed when an HRT of 5 days was applied. Methane productivity reached the maximum value of 1.13 ± 0.08 L/L/d at 10 days HRT whereas the methane yield increased with the HRT. The Anaerobic Digestion Model no. 1 (ADM1) was applied to the obtained experimental data from the methanogenic reactor to simulate the digester response at all HRT tested. The ability of the model to predict the experimental results was evident even in the case of the process failure, thus implying that the ADM1 could be a valuable tool for process design even in the case of a complex feedstock. In general, the two-stage anaerobic digestion proved to be a stable, reliable and effective process for energy recovery and stabilization treatment of olive pulp.     

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.

     

A hybrid anaerobic solid-liquid bioreactor for food waste digestion

A hybrid anaerobic solid-liquid (HASL) bioreactor is an enhanced two-phase anaerobic system, that consists of a solid waste reactor as the acidification reactor and a wastewater reactor, i.e. an upflow anaerobic sludge blanket (UASB) reactor as the methanogenic reactor. Food waste digestion in HASL bioreactors with pre-acidification and HASL operation stages was investigated in two separate runs. After 8 days of pre-acidification in Run A and 4 days in Run B, total volatile fatty acid (TVFA) and chemical oxygen demand (COD) concentrations in the leachates of both acidification reactors were similar. During HASL operation stage, TVFA and COD removal in the methanogenic phase were 77–100% and 75–95%, respectively. Some 99% of the total methane generated was from the methanogenic phase with a content of 68–70% methane. At the end of operation, about 59–60% of the added volatile solids (VS) were removed with a methane yield of 0.25 l g^–1 VS.     

Degradation of cyanide in agroindustrial or industrial wastewater in an acidification reactor or in a single-step methane reactor by bacteria enriched from soil and peels of cassava

During cassava starch production, large amounts of cyanoglycosides were released and hydrolysed by plant-borne enzymes, leading to cyanide concentrations in the wastewater as high as 200 mg/l. For anaerobic degradation of the cyanide during pre-acidification or single-step methane fermentation, anaerobic cultures were enriched from soil residues of cassava roots and sewage sludge. In a pre-acidification reactor this culture was able to remove up to 4 g potassium cyanide/l of wastewater at a hydraulic retention time (t _HR) of 4 days, equivalent to a maximal cyanide space loading of 400 mg CN⁻ l⁻¹ day⁻¹. The residual cyanide concentration was 0.2–0.5 mg/l. Concentrated cell suspensions of the mixed culture formed ammonia and formate in almost equimolar amounts from cyanide. Little formamide was generated by chemical decay. A concentration of up to 100 mmol ammonia/l had no inhibitory effect on cyanide degradation. The optimal pH for cyanide degradation was 6–7.5, the optimal temperature 25–37 °C. At a pH of 5 or lower, cyanide accumulated in the reactor and pre-acidification failed. The minimal t _HR for continuous cyanide removal was 1.5 days. The enriched mixed culture was also able to degrade cyanide in purely mineralic wastewater from metal deburring, either in a pre-acidification reactor with a two-step process or in a one-step methanogenic reactor. It was necessary to supplement the wastewater with a carbon source (e.g. starch) to keep the population active enough to cope with any possible inhibiting effect of cyanide. Received: 29 April 1998 / Received revision: 8 June 1998 / Accepted: 14 June 1998     

Process contribution evaluation for COD removal and energy production from molasses wastewater in a BioH<Subscript>2</Subscript>–BioCH<Subscript>4</Subscript>–MFC-integrated system

In this study, a three-stage-integrated process using the hydrogenic process (BioH₂), methanogenic process (BioCH₄), and a microbial fuel cell (MFC) was operated using molasses wastewater. The contribution of individual processes to chemical oxygen demand (COD) removal and energy production was evaluated. The three-stage integration system was operated at molasses of 20 g-COD L^?1, and each process achieved hydrogen production rate of 1.1 ± 0.24 L-H₂ L^?1 day^?1, methane production rate of 311 ± 18.94 mL-CH₄ L^?1 day^?1, and production rate per electrode surface area of 10.8 ± 1.4 g m^?2 day^?1. The three-stage integration system generated energy production of 32.32 kJ g-COD^?1 and achieved COD removal of 98 %. The contribution of BioH₂, BioCH₄, and the MFC reactor was 20.8, 72.2, and, 7.0 % of the total COD removal, and 18.7, 81.2, and 0.16 % of the total energy production, respectively. The continuous stirred-tank reactor BioH₂ at HRT of 1 day, up-flow anaerobic sludge blanket BioCH₄ at HRT of 2 days, and MFC reactor at HRT of 3 days were decided in 1:2:3 ratios of working volume under hydraulic retention time consideration. This integration system can be applied to various configurations depending on target wastewater inputs, and it is expected to enhance energy recovery and reduce environmental impact of the final effluent.     

Microbial production of a biofuel (acetone–butanol–ethanol) in a continuous bioreactor: impact of bleed and simultaneous product removal

Acetone butanol ethanol (ABE) was produced in an integrated continuous one-stage fermentation and gas stripping product recovery system using Clostridium beijerinckii BA101 and fermentation gases (CO₂ and H₂). In this system, the bioreactor was fed with a concentrated sugar solution (250–500 g L^?1 glucose). The bioreactor was bled semi-continuously to avoid accumulation of inhibitory chemicals and products. The continuous system was operated for 504 h (21 days) after which the fermentation was intentionally terminated. The bioreactor produced 461.3 g ABE from 1,125.0 g total sugar in 1 L culture volume as compared to a control batch process in which 18.4 g ABE was produced from 47.3 g sugar. These results demonstrate that ABE fermentation can be operated in an integrated continuous one-stage fermentation and product recovery system for a long period of time, if butanol and other microbial metabolites in the bioreactor are kept below threshold of toxicity.     

High-Rate two-phase process for the anaerobic degradation of cellulose, employing rumen microorganisms for an efficient acidogenesis

A novel two-stage anaerobic process for the microbial conversion of cellulose into biogas has been developed. In the first phase, a mixed population of rumen bacteria and ciliates was used in the hydrolysis and fermentation of cellulose. The volatile fatty acids (VFA) produced in this acidogenic reactor were subsequently converted into biogas in a UASB-type methanogenic reactor.A stepwise increase of the loading rate from 11.9 to 25.8 g volatile solids/L reactor volume/day (g VS/L/day) did not affect the degradation efficiency in the acidogenic reactor, whereas the methanogenic reactor appeared to be overloaded at the highest loading rate. Cellulose digestion was almost complete at all loading rates applied. The two-stage anaerobic process was also tested with a closed fluid circuit. In this instance total methane production was 0.438 L CH(4)g VS added, which is equivalent to 98% of the theoretical value. The application of rumen microorganisms in combination with a high-rate methane reactor is proposed as a means of efficient anaerobic degradation of cellulosic residues to methane. Because this newly developed two-phase system is based on processes and microorganisms from the ruminant, it will be referred to as "Rumen Derived Anaerobic Digestion" (RUDAD-) process.     

Methane fermentation of coastal mud sediment by a two-stage upflow anaerobic sludge blanket (UASB) reactor system

The removal of organic matter from a coastal mud sediment was carried out by a methane fermentation process under anaerobic conditions. In a batch acidogenic fermentation, the addition of vitamins containing thiamine, nicotinic acid and biotin dramatically enhanced acetate production from the mud sediment (200 g wet wt l(-1) artificial sea water), yielding 77 mM acetate after 6 days, which corresponded to 77% of the organic matter in the mud sediment, measured on the basis of chemical oxygen demand. Thereafter, the two-fold diluted, post-acidogenic fermentation liquor (PAF liquor) was continuously treated at 2.4x original dilution rate day(-1) for 30 days, using an upflow anaerobic sludge blanket methanogenic reactor containing the acclimated methanogenic sludge from the mud sediment. Acetate, 42 mM in the PAF liquor, was converted to methane at a maximum methane production rate of 96 mmol l(-1) day(-1); and 87.5% of the acetate and 88.7% of the total organic carbon in the PAF liquor were removed. Moreover, an efficient treatment of the mud sediment was carried out by a semi-continuous, two-stage reactor system, where the culture broth was circulated between acidogenic and methanogenic reactors. This two-stage reactor system gave a stable operation at 4-day intervals for one treatment period, yielding 112 mmol methane from the wet mud in the PAF liquor (278 g l(-1)).     

Effect of low pH start-up on continuous mixed-culture lactic acid fermentation of dairy effluent

Mixed-culture fermentation that does not require an energy-intensive sterilization process is a viable approach for the economically feasible production of lactic acid (LA) due to the potential use of organic waste as feedstock. This study investigated mixed-culture LA fermentation of whey, a high-strength organic wastewater, in continuous mode. Variations in the hydraulic retention time (HRT) from 120 to 8 h under different pH regimes in two thermophilic reactors (55 °C) were compared for their fermentation performance. One reactor was maintained at a low pH (pH 3.0) during operation at HRTs of 120 to 24 h and then adjusted to pH 5.5 in the later phases of fermentation at HRTs of 24 to 8 h (R1), while the second reactor was maintained at pH 5.5 throughout the experiment (R2). Although the LA production in R1 was negligible at low pH, it increased dramatically after the pH was raised to 5.5 and exceeded that in R2 when stabilized at HRTs of 8 and 12 h. The maximum yield (0.62 g LA/g substrate fed as the chemical oxygen demand (COD) equivalent), the production rate (11.5 g/L day), and the selectivity (95 %) of LA were all determined at a 12-h HRT in R1. Additionally, molecular and statistical analyses revealed that changes in the HRT and the pH significantly affected the bacterial community structure and thus the fermentation characteristics of the experimental reactors. Bacillus coagulans was likely the predominant LA producer in both reactors. The overall results suggest that low pH start-up has a positive effect on yield and selectivity in mixed-culture LA fermentation.     

Application of a fuzzy logic control system for continuous anaerobic digestion of low buffered, acidic energy crops as mono-substrate

A fuzzy logic control (FLC) system was developed at the Hamburg University of Applied Sciences (HAW Hamburg) for operation of biogas reactors running on energy crops. Three commercially available measuring parameters, namely pH, the methane (CH4) content, and the specific gas production rate (spec. GPR = m(3)/kg VS/day) were included. The objective was to avoid stabilization of pH with use of buffering supplements, like lime or manure. The developed FLC system can cover most of all applications, such as a careful start-up process and a gentle recovery strategy after a severe reactor failure, also enabling a process with a high organic loading rate (OLR) and a low hydraulic retention time (HRT), that is, a high throughput anaerobic digestion process with a stable pH and CH4 content. A precondition for a high load process was the concept of interval feeding, for example, with 8 h of interval. The FLC system was proved to be reliable during the long term fermentation studies over 3 years in one-stage, completely stirred tank reactors (CSTR) with acidic beet silage as mono-input (pH 3.3-3.4). During fermentation of the fodder beet silage (FBS), a stable HRT of 6.0 days with an OLR of up to 15 kg VS/m(3)/day and a volumetric GPR of 9 m(3)/m(3)/day could be reached. The FLC enabled an automatic recovery of the digester after two induced severe reactor failures. In another attempt to prove the feasibility of the FLC, substrate FBS was changed to sugar beet silage (SBS), which had a substantially lower buffering capacity than that of the FBS. With SBS, the FLC accomplished a stable fermentation at a pH level between 6.5 and 6.6, and a volatile fatty acid level (VFA) below 500 mg/L, but the FLC had to interact and to change the substrate dosage permanently. In a further experiment, the reactor temperature was increased from 41 to 50 degrees C. Concomitantly, the specific GPR, pH and CH4 dropped down. Finally, the FLC automatically enabled a complete recovery in 16 days.     

Mesophilic biogas production from fruit and vegetable waste in a tubular digester

A semi-continuously mixed mesophilic tubular anaerobic digester was tested for the conversion of fruit and vegetable waste (FVW) into biogas. The effect of hydraulic retention time (HRT) and the feed concentration on the extent of the degradation of the waste was examined. Varying the HRT between 12 and 20 days had no effect on the fermentation stability and pH remained between 6.8 and 7.6, but an inhibition of methanogenic bacteria was observed at HRT below 12 days. The overall performance of the reactor was depressed by changing the feed concentration from 8% to 10% TS (dry weight). By applying a feed concentration of 6% and HRT of 20 days in the tubular digester, 75% conversion efficiency of FVW into biogas with a methane content of 64% was achieved.     

High rate treatment of terephthalic acid production wastewater in a two-stage anaerobic bioreactor

The feasibility was studied of anaerobic treatment of wastewater generated during purified terephthalic acid (PTA) production in two-stage upflow anaerobic sludge blanket (UASB) reactor system. The artificial influent of the system contained the main organic substrates of PTA-wastewater: acetate, benzoate, and terephthalate. Three parallel operated reactors were used for the second stage, and seeded with a suspended terephthalate degrading culture, with and without additional methanogenic granular sludge (two different types). The first stage UASB-reactor was seeded with methanogenic granular sludge. Reactors were operated at 37 degrees C and pH 7. During the first 300 days of operation a clear distinction between the biomass grown in both reactor stages was obtained. In the first stage, acetate and benzoate were degraded at a volumetric loading rate of 40 g-COD/L . day at a COD-removal efficiency of 95% within the first 25 days of operation. No degradation of terephthalate was obtained in the first stage during the first 300 days of operation despite operation of the reactor at a decreased volumetric loading rate with acetate and benzoate of 9 g-COD/L . day from day 150. Batch incubation of biomass from the reactor with terephthalate showed that the lag-phase prior to terephthalate degradation remained largely unchanged, indicating that no net growth of terephthalate degrading biomass occurred in the first stage reactor. From day 300, however, terephthalate degradation was observed in the first stage, and the biomass in this reactor could successfully be enriched with terephthalate degrading biomass, resulting in terephthalate removal capacities of 15 g-COD/L . day. Even though no single reason could be identified why (suddenly) terephthalate degradation was obtained after such a long period of operation, it is suggested that the solid retention time as well the prevailing reactor concentrations acetate and benzoate may have played an important role. From day 1 of operation, terephthalate was degraded in the second stage. In presence of methanogenic granular biomass, high terephthalate removal capacities were obtained in these reactors (15 g-COD/L . day) after approximately 125 days of operation. From the results obtained it is concluded that terephthalate degradation is the bottleneck during anaerobic treatment of PTA-wastewater. Pre-removal of acetate and benzoate in staged bioreactor reduces the lag-phase prior to terephthalate degradation in latter stages, and enables high rate treatment of PTA-wastewater.     

Analysis of the methane production in thermophilic anaerobic reactors: use of autofluorescence microscopy

Methanogenic activity in thermophilic, anaerobic reactors was determined by comparing the amount of methane generated in single- and two-stage systems with the size of the methanogenic population, as determined by microscopy. The methanogenic activities were 2.71 × 10^–9 ml methane cell^–1 d^–1 and 1.10 × 10^–9 ml methane cell^–1 d^–1 for 10 and 4 days of the hydraulic retention time (HRT), in the single-stage system. In the two-stage system, 7.49 × 10^–9 ml methane cell^–1 d^–1 in the acidogenic reactor and 1.56 × 10^–9 ml methane cell^–1 d^–1 in the methanogenic reactor for 4 days of the HRT. A high correlation was evident between the methane production and methanogenic population [0.1354 ln(x) – 2.1375](R ² 0.8619).     

Mesophilic and thermophilic aerobic digestion of municipal sludge in an airlift U-shape bioreactor

Aerobic digestion of primary and secondary sludges was studied in airlift bioreactors at mesophilic and thermophilic temperatures. The experimental studies were conducted with a laboratory U-shape airlift reactor (operating volume 23 liters) and in a pilot U-shape airlift reactor of 1150 liters operating volume. In the laboratory reactor, with cold (6°C) and concentrated (3–4% solids) feed of primary and secondary municipal sludge, a 30% volatile suspended solids (VSS) reduction was achieved with a hydraulic retention time (HRT) of 2·5 days. A VSS loading rate of 8·2 kg VSS/m³/day was achieved. This loading is comparable to that obtained in a pure-oxygen sparged, mixed reactor.In the pilot-plant reactor at mesophilic temperature (31–33°C), a VSS loading rate of 7·9 kg VSS/m³/day and a VSS reduction of 40% were achieved with a HRT of 4 days.