Ammonia–methane two-stage anaerobic digestion of dehydrated waste-activated sludge

The study investigated methane production from dehydrated waste-activated sludge (DWAS) with approximately 80% water content under thermophilic conditions. The repeated batch-wise treatment of DWAS using methanogenic sludge unacclimated to high concentrations of ammonia, increased the ammonia production up to 7,600 mg N per kilogram total wet sludge of total ammonia concentration, and stopped the methane production. Investigation revealed that the loading ratio of DWAS for methanogenic sludge influences anaerobic digestion. Methane production significantly decreased and ammonia concentration increased with the increase in loading ratio of DWAS. Since the semicontinuous culture revealed that approximately 50% of organic nitrogen in DWAS converted to ammonia at sludge retention time (SRT) after 4 days at 37 degrees C and 1.33 days at 55 degrees C, the previous stripping of the ammonia produced from DWAS was carried out. The stripping of ammonia increased methane production significantly. This ammonia-methane two-stage anaerobic digestion demonstrated a successful methane production at SRT 20 days in the semicontinuous operation using a laboratory-scale reactor system.     

Dry anaerobic ammonia–methane production from chicken manure

The effect of temperature on production of ammonia during dry anaerobic fermentation of chicken manure (CM), inoculated with thermophilic methanogenic sludge, was investigated in a batch condition for 8 days. Incubation temperature did not have a significant effect on the production of ammonia. Almost complete inhibition of production of methane occurred at 55 and 65°C while quite low yields of 8.45 and 6.34 ml g⁻¹ VS (volatile solids) were observed at 35 and 45°C due to a higher accumulation of ammonia. In order to improve the production of methane during dry anaerobic digestion of CM, stripping of ammonia was performed firstly on the CM previously fermented at 65°C for 8 days: the stripping for 1 day at 85°C and pH 10 removed 85.5% of ammonia. The first-batch fermentation of methane for 75 days was conducted next, using the ammonia-stripped CM inoculated with methanogenic sludge at different ratios, (CM: thermophilic sludge) of 1:2, 1:1, and 2:1 on volume per volume basis at both 35 and 55°C. Production of methane improved and was higher than that of the control (without stripping of ammonia) but the yield of 20.4 ml g⁻¹ VS was still low, so second stripping of ammonia was conducted, which resulted in 74.7% removal of ammonia. A great improvement in the production of methane of 103.5 ml g⁻¹ VS was achieved during the second batch for 55 days.     

Performance of an UASB reactor treating synthetic wastewater at low-temperature using cold-adapted seed slurry

The present study is an attempt to investigate if a long-term acclimation of digester contents to low-temperatures would improve wastewater treatment at low-temperatures similar to mesophilic ranges. The feasibility of low-temperature (15 °C) anaerobic treatment of synthetic wastewater in an upflow anaerobic sludge blanket reactor was studied using inoculum from a cattle manure digester adapted to 15 °C. The effect of varying hydraulic retention time was studied by decreasing the retention time from 7 days to 1 day. Under a constant temperature of 15 °C with a hydraulic retention time of 1 day and a corresponding loading rate of 7.2 g-chemical oxygen demand (COD)/l/day, 90–95% removal efficiency was achieved. The methane production of 250 l/kg-COD removed at standard temperature pressure (STP) is a major highlight of the study complementing the high treatment efficiency achieved. Loading rates >5 g-COD/l/day was accompanied by increase in effluent volatile fatty acids (VFA) concentrations. Due to the presence of a high concentration of active granular sludge in the lower compartment of the reactor, 80% reduction of COD occurred within the granular bed of the reactor. Treatment of low strength wastewater for a short period showed 70–75% removal efficiencies with methane yield of 300 l/kg-COD removed. Specific methanogenic activity profiles of the anaerobic biomass revealed low-temperature (15 °C) optima, indicating selection of cold-active microorganisms during the acclimation process. The SMA assays also indicate the development of a putatively psychrophilic acetoclastic methanogenic community and biogas analysis showed 75% efficiency in energy recovery as methane.     

Factors in the determination of methanogenic potential of manure

The influence of the substrate concentration, the micro and macro nutrients and buffer requirements, the sludge origin (biomass that is acclimatized or not acclimatized to waste) and the inoculum/substrate ratio (ISR) were studied to determine their effects in the methanogenic potential of turkey manure, which is a solid waste. According to the results obtained, the methane production determination does not require the addition of nutrients (additional to the contents in the waste) and a buffer for this type of assay. The methane yield (Y_CH4) performance is given by the substrate concentration and the sludge origin, therefore it is better to carry out the assay with biomass that is already adapted to the waste. The methanogenic potential of this type of waste is not determined by the amount of sludge and it does not need an external inoculum (external to the waste contents).     

Nitrogen removal from wastewater with a low COD/N ratio at a low oxygen concentration

The goal of the study was to determine the effectiveness of nitrification and denitrification and the kinetics of ammonia removal from a mixture of wastewater and anaerobic sludge digester supernatant in an SBR at limited oxygen concentration. In addition, the COD removal efficiency and sludge production were assessed.In the SBR cycle alternating aerobic and anaerobic phases occurred; in the aeration phase the dissolved oxygen (DO) concentration was below 0.7 mg O₂/L. The low DO concentration did not inhibit ammonia oxidation-nitrification and the efficiency was ca. 96-98%. However, a relatively high COD concentration in the effluent was detected. The values of K_m and V_max, calculated from the Michaelis-Menten equation, were 43 mg N-NH₄/L and 15.64 mg N-NH₄/L h, respectively. Activated sludge production was almost stable (0.62-0.66 g MLVSS/g COD). A high net biomass production resulted from a low specific biomass decay rate of 0.0015 d⁻¹.     

Effect of ammonia on the methanogenic activity of methylaminotrophic methane producing Archaea enriched biofilm

Ammonia is a metabolic product in the decomposition of protein wastes, and has a recognized inhibitory effect on methanogenesis; this effect has been slightly quantified on methanogenic biofilms and particularly those populated by methanogenic Archaea which produce ammonia as a catabolic product from methylated amines. This paper presents studies on the effect of ammonia on maximum methanogenic activity of anaerobic biofilms enriched by methylaminotrophic methane producing Archaea (mMPA). The effect of unionized free ammonia on the specific maximum methanogenic activity of a mMPA enriched biofilm was studied, using 250 mL flasks containing ceramic rings colonized by 30 day-old experimental biofilm and adding 48.8 (control system), 73.8, 98.8, 148.8, 248.8, 448.8 and 848.8 mg NH(3)-N/L. The systems were maintained for ten days at a pH of 7.5 and temperature of 37 degrees C. The results showed that at 848.8 mg NH(3)-N/L, biofilm methane production required 36 h adaptation period, prior to entering into maximum production phase. The highest maximum methanogenic activity reached a value of 2.337+/-0.213 g COD methane/g VSS *day when 48.8 mg NH(3)-N/L was added, and inhibition was clearly observed in those systems above 148.8 mg NH(3)-N/L, producing under 1.658+/-0.185 g COD methane/g VSS *day. The lowest methanogenic activity reached was 0.639+/-0.162 g COD methane/g VSS *day at the system added with 848.8 mg NH(3)-N/L. When applying the Luong and non-competitive inhibition models, the best fit was obtained with the non-competitive model, which predicted 50% inhibition of methanogenic activity at 365.288 mg NH(3)-N/L.     

The roles of acetotrophic and hydrogenotrophic methanogens during anaerobic conversion of biomass to methane: a review

Among different conversion processes for biomass, biological anaerobic digestion is one of the most economic ways to produce biogas from various biomass substrates. In addition to hydrolysis of polymeric substances, the activity and performance of the methanogenic bacteria is of paramount importance during methanogenesis. The aim of this paper is primarily to review the recent literature about the occurrence of both acetotrophic and hydrogenotrophic methanogens during anaerobic conversion of particulate biomass to methane (not wastewater treatment), while this review does not cover the activity of the acetate oxidizing bacteria. Both acetotrophic and hydrogenotrophic methanogens are essential for the last step of methanogenesis, but the reports about their roles during this phase of the process are very limited. Despite, some conclusions can still be drawn. At low concentrations of acetate, normally filamentous Methanosaeta species dominate, e.g., often observed in sewage sludge. Apparently, high concentrations of toxic ionic agents, like ammonia, hydrogen sulfide (H₂S) and volatile fatty acids (VFA), inhibit preferably Methanosaetaceae and especially allow the growth of Methanosarcina species consisting of irregular cell clumps, e.g., in cattle manure. Thermophilic conditions can favour rod like or coccoid hydrogenotrophic methanogens. Thermophilic Methanosarcina species were also observed, but not thermophilic Methanosaetae. Other environmental factors could favour hydrogentrophic bacteria, e.g., short or low retention times in a biomass reactor. However, no general rules regarding process parameters could be derivated at the moment, which favours hydrogenotrophic methanogens. Presumably, it depends only on the hydrogen concentration, which is generally not mentioned in the literature.     

Acidophilic degradation of methanol by a methanogenic enrichment culture

Abstract An acidophilic methanogenic enrichment culture was obtained in a continuous up-flow anaerobic sludge blanket reactor operated at pH 4.2 with methanol as the sole carbon source. The specific methylotrophic methanogenic activity of the enriched reactor sludge at pH 5 was 3.57 g COD g⁻¹ volatile suspended solids day⁻¹ and the apparent doubling time of the biomass was 15.8 h. Acidic conditions were obligatory, since the enrichment culture was not able to produce methane or to grow at pH 7. Based on morphological characteristics, the dominant methanogenic species in the enrichment culture was a Methanosarcina .     

Enhancement of biochemical methane potential from excess sludge with low organic content by mild thermal pretreatment

Excess sludge with low organic content always led to the failure of anaerobic digestion for methane production. Recently, the mild thermal pretreatment, which is usually operated at temperatures below 120 °C, has drawn much attention due to less energy consumption and no chemical addition. In this study the effect of mild thermal pretreatment (50–120 °C) on the solubilization and methane potential of excess sludge with a low concentration of organic matters was investigated. Experimental results showed that the concentration of soluble organic matters increased gradually with temperature during the mild thermal pretreatment of excess sludge. Biochemical methane potential experiments demonstrated that the potential of methane production from excess sludge was greatly enhanced by mild thermal pretreatment, and under the conditions of pretreatment temperature 100 °C and digestion time 20 d the methane yield was as high as 142.6 ± 2.5 mL/g of volatile solids. Mechanism investigation on the enhancement of methane production from excess sludge exhibited that the consumptions of sludge protein and carbohydrate, the adenosine 5′-triphosphate content of anaerobic microorganisms, the activities of key enzymes related to anaerobic digestion, and the amount of methanogens were all improved by mild thermal pretreatment, in correspondence with the production of methane.     

Promotion of polylactide degradation by ammonia under hyperthermophilic anaerobic conditions

The objective of this study was to evaluate the promotion effect of ammonia on the biodegradation of polylactide (PLA) under hyperthermophilic (80 °C) and thermophilic (55 °C) anaerobic condition. The results showed that PLA was transformed to lactic acid under hyperthermophilic conditions, but that the transformation ratio was negligible under thermophilic conditions. The hydrolysis process can be markedly increased with ammonia addition and microorganism activity. The maximum transformation ratios of the two kinds of PLA used in this study were 65.2% and 51.8%, respectively, with ammonia addition of 4 g N/L over 3 days treatment of anaerobic sludge. After the hyperthermophilic pretreatment, the hydrolysis products were converted to methane by methanogens under the thermophilic and anaerobic conditions. The final methane conversion ratios of the two kinds of PLA after 22 days treatment were 81.8% and 77.0%, respectively.     

Start-up and inhibition analysis of the Anammox process seeded with anaerobic granular sludge

The longer start-up period of the Anammox process is due to the very low cellular yield and growth rates of Anammox bacteria. Nitrite inhibition is considered to be the key factor in the instability of the Anammox process during the operation. However, little attention was paid to the inhibitory effect of pH and free ammonia. This paper presents start-up and inhibition analysis of an Anammox biofilm reactor seeded with anaerobic granular sludge. Results showed that the start-up period could be divided into the sludge lysis phase, lag phase, propagation phase, stationary phase and inhibition phase. Optimization control could be implemented correspondingly to accelerate the start-up of Anammox bioreactors. Effluent pH increased to 8.7–9.1 when the nitrogen removal rate was higher than 1,200 mg l⁻¹ day⁻¹. The free ammonia concentration was accompanied with a higher level of 64–73 mg l⁻¹. Inhibitory effects of high pH and free ammonia on Anammox bacteria contributed to the destabilization of the Anammox bioreactor during the first 125 days with influent KHCO₃ of 0.5 g l⁻¹. Increasing the suffering capacity in the inlet by dosing 1.25 g KHCO₃ l⁻¹ effectively reduced the pH variation, and the nitrogen removal performance of the reactor was further developed.     

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.     

Effects of the acidogenic biomass on the performance of an anaerobic membrane bioreactor for wastewater treatment

Continuous flow experiments were performed to study the effects of acidogenic biomass development, induced by feeding with non-acidified substrate, on the operation and performance of an anaerobic membrane bioreactor (AnMBR). The AnMBR was operated at cross-flow velocities up to 1.5 m/s and fed with a gelatine–starch–ethanol mixture. A significant fraction of acidogenic biomass developed during reactor operation, which fully determined the sludge rheology, and influenced the particle size distribution. As a result, flux levels of only 6.5 l/m² h were achieved, at a liquid superficial velocity of 1.5 m/s. Even though the soluble microbial products levels in the AMBR were as high as 14 g COD/l, the observed hydraulic flux was not limited by irreversible pore fouling, but by reversible cake layer formation. Propionate oxidation was the limiting step for the applied organic loading rate. The assessed specific methanogenic activity (SMA) with propionate as substrate was, however, similar to the values found by others during thermophilic treatment of non or partially acidified substrates in granular sludge bed reactors, indicating an appropriate level of the propionate oxidation capacity.     

Nitrogen removal performance of a hybrid anammox reactor

The anaerobic ammonium oxidation (anammox) process has attracted considerable attention in recent years as an alternative to conventional nitrogen removal technologies. In this study, an innovative hybrid reactor combining fluidized and fixed beds for anammox treatment was developed. The fluidized bed was mechanically stirred and the gaseous product could be rapidly released from the anammox sludge to prevent washout of the sludge caused by floatation. The fixed bed comprising a non-woven biomass carrier could efficiently catch sludge to reduce washout. During the operation, nitrogen loading rates to the reactor were increased to 27.3 kg N/m³/d, with total nitrogen removal efficiencies of 75%. The biomass concentration in the fluidized bed reached 26-g VSS/L. Anammox granules were observed in the reactors, with settling velocities and sludge volumetric index of 27.3 ± 6.5 m/h and 23 mL/g, respectively. Quantification of extracellular polymeric substances revealed the anammox granules contained a significant amount of extracellular proteins.     

Decreasing ammonia inhibition in thermophilic methanogenic bioreactors using carbon fiber textiles

Ammonia accumulation is one of the main causes of the loss of methane production observed during fermentation. We investigated the effect of addition of carbon fiber textiles (CFT) to thermophilic methanogenic bioreactors with respect to ammonia tolerance during the process of degradation of artificial garbage slurry, by comparing the performance of the reactors containing CFT with the performance of reactors without CFT. Under total ammonia-N concentrations of 3,000 mg L⁻¹, the reactors containing CFT were found to mediate stable removal of organic compounds and methane production. Under these conditions, high levels of methanogenic archaea were retained at the CFT, as determined by 16S rRNA gene analysis for methanogenic archaea. In addition, Methanobacterium sp. was found to be dominant in the suspended fraction, and Methanosarcina sp. was dominant in the retained fraction of the reactors with CFT. However, the reactors without CFT had lower rates of removal of organic compounds and production of methane under total ammonia-N concentrations of 1,500 mg L⁻¹. Under this ammonia concentration, a significant accumulation of acetate was observed in the reactors without CFT (130.0 mM), relative to the reactors with CFT (4.2 mM). Only Methanobacterium sp. was identified in the reactors without CFT. These results suggest that CFT enables stable proliferation of aceticlastic methanogens by preventing ammonia inhibition. This improves the process of stable garbage degradation and production of methane in thermophilic bioreactors that include high levels of ammonia.     

Effect of pH on phase separated anaerobic digestion

A pilot scale experiment was performed for a year to develop a two-phase anaerobic process for piggery wastewater treatment (COD: 6,000 mg/L, BOD: 4,000 mg/L, SS: 500 gm/L, pH 8.4, alkalinity 6,000 mg/L). The acidogenic reactor had a total volume of 3 m³, and the methanogenic reactor, an, anaerobic up-flow sludge filter, combining a filter and a sludge bed, was also of total volume 3 m³ (1.5 m³ of upper packing material). Temperatures of the acidogenic and methanogenic reactors kept at 20°C and 35°C., respectively. When the pH of the acidogenic reactor was controlled at 6.0–7.0 with HCl, the COD removal efficiency increased from 50 to 80% over a period of six months, and as a result, the COD of the final effluent fell in the range of 1,000–1,500 mg/L. BOD removal efficiency over the same period was above 90%, and 300 to 400 mg/L was maintained in the final effluent. The average SS in the final effluent was 270 mg/L. The methane production was 0.32 m³ CH₄/kg COD_removed and methane content of the methanogenic reactor was high value at 80–90%., When the pH of the acidogenic reactor was not controlled over the final two months, the pH reached 8.2 and acid conversion decreased compared with that of pH controlled, while COD removal was similar to the pH controlled operation. Without pH control, the methane content in the gas from methanogenic reactor improved to 90%, compared to 80% with pH control.     

Anaerobic waste-activated sludge digestion-a bioconversion mechanism and kinetic model

The anaerobic bioconversion of raw and mechanically lysed waste-activated sludge was kinetically investigated. The hydrolysis of the biopolymers, such as protein, which leaked out from the biological sludge with ultrasonic lysis, was a first-order reaction in anaerobic digestion and the rate constant was much higher that the decay rate constant of the raw waste activated sludge. An anaerobic digestion model that is capable of evaluating the effect of the mechanical sludge lysis on digestive performance was developed. The present model includes four major biological processes-the release of intracellular matter with sludge lysis; hydrolysis of biopolymers to volatile acids; the degradation of various volatile acids to acetate; and the conversion of acetate and hydrogen to methane. Each process was assumed to follow first order kinetics. The model suggested that when the lysed waste-activated sludge was fed, the overall digestive performance remarkably increased in the two-phase system consisting of an acid forming process and a methanogenic process, which ensured the symbiotic growth of acetogenic and methanogenic bacteria. (c) 1993 Wiley & Sons, Inc.     

The effect of low temperature (5–29 °C) and adaptation on the methanogenic activity of biomass

The influence of low temperature (5–29 °C) on the methanogenic activity of non-adapted digested sewage sludge and on temperature/leachate-adapted biomass was assayed by using municipal landfill leachate, intermediates of anaerobic degradation (propionate) and methane precursors (acetate, H₂/CO₂) as substrates. The temperature dependence of methanogenic activity could be described by Arrhenius-derived models. However, both substrate and adaptation affected the temperature dependence. The adaptation of biomass in a leachate-fed upflow anaerobic sludge-blanket reactor at approximately 20 °C for 4 months resulted in a sevenfold and fivefold increase of methanogenic activity at 11 °C and 22 °C respectively. Both acetate and H₂/CO₂ were methanized even at 5 °C. At 22 °C, methanogenic activities (acetate 4.8–84 mM) were 1.6–5.2 times higher than those at 11 °C. The half-velocity constant (K _s) of acetate utilization at 11 °C was one-third of that at 22 °C while a similar K _i was obtained at both temperatures. With propionate (1.1–5.5 mM) as substrate, meth‐anogenic activities at 11 °C were half those at 22 °C. Furthermore, the residual concentration of the substrates was not dependent on temperature. The results suggest that the adaptation of biomass enables the achievement of a high treatment capacity in the anaerobic process even under psychrophilic conditions. Received: 23 December 1996 / Received last revision: 18 June 1997 / Accepted: 23 June 1997     

Methanogenic toxicity in anaerobic digesters treating municipal wastewater

In upflow anaerobic sludge blanket (UASB) digesters treating raw sewage at low temperatures, the sludge progressively lost methanogenic activity, indicating the possibility of methanogenic activity inhibition caused by wastewater constituents. To check this fact, batch and semi-continuous methanogenic toxicity assays were carried out with raw and centrifuged sewage. Permanent methanogenic toxicity on anaerobic sludge of approximately 50% was found when the sludge exposure to wastewater was renewed in a semi-continuous way. A stronger methanogenic inhibition of about 70-100% was observed when an active anaerobic sludge was exposed to mixed liquor from the UASB digester treating municipal wastewater. Suspended solids removal from sewage slightly reduced methanogenic toxicity. Effective concentration of municipal wastewater that caused a 50% reduction in methanogenic activity was estimated to be in the range of 150-200 mg CODl(-1). As methanogenic inhibition appeared to be related to remaining COD, higher methanogenic toxicity in digesters operating with low conversion efficiency will be expected.