Potential of biogas recirculation to enhance biomass accumulation on supporting media

Two lab-scale anaerobic hybrid reactors (AHR) were operated to investigate the effect of recirculated biogas on the development of biomass on supporting media during the start-up. The reactor comprised of two distinct zones; sludge bed on the bottom and packed bed using nylon fiber as the media on the upper half of the reactor. Both reactors were continuously fed with cassava starch wastewater. The organic loading rate (OLR) was increased from 0.3 to 5.5 g COD/L/day by gradually decreasing the hydraulic retention time (HRT) from 37 to 3.5 days in two months. The biogas at 2.6 L/L/day was recirculated merely in the first month of the operation in order to allow the attached biomass to grow according to the organic matters present in the reactor at the final stage of the start up. Chemical oxygen demand (COD) removal efficiency of over 80% was achieved throughout the study. The result demonstrated a better COD removal efficiency for the reactor with biogas recirculation, especially at low HRTs. The amounts of biomass accumulated on the media in both reactors were slightly different with 11.9 gVSS found on the one with biogas recirculation compared to 9.8 gVSS on the other. In addition, 16.3% increase of the sludge bed was achieved with biogas recirculation as opposed to 9% in the control one. The attached biomass activity test indicated a greater amount and more favorable ratio of the methanogenic bacterial group on the media with the recirculation correlating well to a relatively higher methane content in biogas. As a result, the recirculation of biogas has a potential of improving the characteristics of the AHR especially in terms of biomass accumulation.     

Research on soybean protein wastewater treatment by the integrated two-phase anaerobic reactor

The start-up tests of treating soybean protein wastewater by the integrated two-phase anaerobic reactor were studied. The results showed that the soybean protein wastewater could be successfully processed around 30 days when running under the situation of dosing seed sludge with the influent of approximately 2000 mg/L and an HRT of 40 h. When the start-up was finished, the removal rate of COD by the reactor was about 80%. In the zone I, biogas mainly revealed carbon dioxide (CO₂) and hydrogen (H₂). Methane was the main component in the zone 2 which ranged from 53% to 59% with an average of 55%. The methane content in biogas increased from the zone I to II. It indicated that the methane-producing capacity of the anaerobic sludge increased. It was found that the uniquely designed two-phase integrated anaerobic reactor played a key role in treating soybean protein wastewater. The acidogenic fermentation bacteria dominated in the zone I, while methanogen became dominant in the zone II. It realized the relatively effective separation of hydrolysis acidification and methanogenesis process in the reactor, which was benefit to promote a more reasonable space distribution of the microbial communities in the reactor. There were some differences between the activities of the sludge in the two reaction zones of the integrated two-phase anaerobic reactor. The activity of protease was higher in the reaction zone I. And the coenzyme F₄₂₀ in the reaction zone II was twice than that in the reaction zone I, which indicated that the activity of the methanogens was stronger in the reaction zone II.     

Searching for links in the biotic characteristics and abiotic parameters of nine different biogas plants

To find links between the biotic characteristics and abiotic process parameters in anaerobic digestion systems, the microbial communities of nine full‐scale biogas plants in South Tyrol (Italy) and Vorarlberg (Austria) were investigated using molecular techniques and the physical and chemical properties were monitored. DNA from sludge samples was subjected to microarray hybridization with the ANAEROCHIP microarray and results indicated that sludge samples grouped into two main clusters, dominated either by Methanosarcina or by Methanosaeta, both aceticlastic methanogens. Hydrogenotrophic methanogens were hardly detected or if detected, gave low hybridization signals. Results obtained using denaturing gradient gel electrophoresis (DGGE) supported the findings of microarray hybridization. Real‐time PCR targeting Methanosarcina and Methanosaeta was conducted to provide quantitative data on the dominating methanogens. Correlation analysis to determine any links between the microbial communities found by microarray analysis, and the physicochemical parameters investigated was conducted. It was shown that the sludge samples dominated by the genus Methanosarcina were positively correlated with higher concentrations of acetate, whereas sludge samples dominated by representatives of the genus Methanosaeta had lower acetate concentrations. No other correlations between biotic characteristics and abiotic parameters were found. Methanogenic communities in each reactor were highly stable and resilient over the whole year.     

Effective anaerobic treatment of fresh leachate from MSW incineration plant and dynamic characteristics of microbial community in granular sludge

We investigated the treatment of fresh leachate from municipal solid waste incineration plants with high-strength organics using a lab-scale expanded granular sludge bed (EGSB) reactor. The reactor was operated at a mesophilic temperature (33 °C) for 118 days. The influent chemical oxygen demand (COD) of the leachate gradually increased to over 70,000 mg/L, and the organic loading rate increased to 18 kg COD/(m³?day). An average COD removal efficiency of 86.7 % was achieved when the reactor was fed with raw leachate, which suggests the feasibility of the EGSB process for leachate treatment. The microbial communities in the sludge from the reactor during the trial operation were constructed by denaturing gradient gel electrophoresis, clone libraries, and real-time quantitative polymerase chain reaction. The dominant group for archaea was Methanosaeta, with 68.4 % proportion at the start of the operation, and then changed to Methanosarcina, with a proportion of 62.3 %, after 118 days of operation. The dominant group of eubacteria was confirmed to be Firmicutes throughout the operation process, with the proportion increasing from >50 to 81.2 %. Almost all the operational taxonomic units of Firmicutes belonged to the order Clostridiales, with characteristic spore formation. The microbial diversity of the population was low under raw leachate as feed in the reactor. The dynamics of the microbial community in the anaerobic granular sludge was discussed relating with the operating status of the EGSB reactor.     

The influence of variable organic loading on the start-up of an anaerobic fluidised bed reactor

Summary A stepped-loading start-up regime utilising variable organic influent concentrations in the range 1650–11600 mgCOD1^–1 was applied to an anaerobic fluidised bed bioreactor at 37°C. The reactor was sensitive to variable influent COD concentrations, but the stepped-loading aided rapid recovery from transient organic loading shocks. Variable effluent COD levels were produced but a COD removal efficiency of 76% was obtained at a final HRT of 0.5 d and an organic loading rate of 5.3 kg COD m^–3 d^–1.     

Methane production from acetamide in an upflow anaerobic sludge-blanket reactor based on a synergistic association between an aerobic rod and methanogens

Acetamide degradation was investigated in a bench-scale upflow anaerobic sludge-blanket (UASB) reactor, successively fed with acetamide, acetate and acetamide, over a period of 343 days, at different hydraulic retention times (t _HR). The reactor was seeded with the sludge previously described [Guyot et al. (1994) Appl Microbiol Biotechnol, 42:452-456], in which methanogenesis from acetamide was performed through a synergistic relationship between an acetamide-degrading, aerobic rod and methanogens. When the reactor was fed acetamide, the chemical oxygen demand (COD) removal efficiency was 86% at volumetric loads less than 1.18 kg COD m^–3 day ^–1. At higher volumetric loads, the efficiency decreased markedly, e.g. 50.9% at a volumetric organic load of 3.39 kg COD m^–3 day^–1 (1 day t _HR) with an accumulation of both acetamide and acetate. The same reactor, when fed with acetate at t _HR 1 day, reached a high COD removal (99%). Evidence of the inhibition of acetate degradation by acetamide is presented. After a long period (135 days) without feeding the reactor with acetamide, the sludge reactor was still capable of degrading acetamide when this substrate was supplied again. It seems that the synergistic degradation of acetamide by aerobes and methanogens present in the UASB reactor sludge is stable over a long period (343 days), in spite of limiting concentrations of dissolved oxygen in the feed.     

Effect of starch addition on the biological conversion and microbial community in a methanol-fed UASB reactor during long-term continuous operation

The effect of starch addition on the microbial composition and the biological conversion was investigated using two upflow anaerobic sludge bracket (UASB) reactors treating methanolic wastewater: one reactor was operated with starch addition, and another reactor was operated without starch addition. Approximately 300 days of operation were performed at 30 kg COD/m³/d, and then, the organic load of the reactors was gradually increased to 120 kg COD/m³/d. Successful operation was achieved at 30 kg COD/m³/d in both reactors; however, the methanol-fed reactor did not perform well at 120 kg COD/m³/d while the methanol-starch-fed reactor did. The granule analysis revealed the granule developed further only in the methanol-starch-fed reactor. The results of the microbial community analysis revealed more Methanosaeta cells were present in the methanol-starch-fed reactor, suggesting the degradation of starch produced acetate as an intermediate, which stimulated the growth of Methanosaeta cells responsible for the extension of granules.     

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.     

Endurance of methanogenic archaea in anaerobic bioreactors treating oleate-based wastewater

Methanogenic archaea are reported as very sensitive to lipids and long chain fatty acids (LCFA). Therefore, in conventional anaerobic processes, methane recovery during LCFA-rich wastewater treatment is usually low. By applying a start-up strategy, based on a sequence of step feeding and reaction cycles, an oleate-rich wastewater was efficiently treated at an organic loading rate of 21 kg COD m^?3 day^?1 (50 % as oleate), showing a methane recovery of 72 %. In the present work, the archaeal community developed in that reactor is investigated using a 16S rRNA gene approach. This is the first time that methanogens present in a bioreactor converting efficiently high loads of LCFA to methane are monitored. Denaturing gradient gel electrophoresis profiling showed that major changes on the archaeal community took place during the bioreactor start-up, where phases of continuous feeding were alternated with batch phases. After the start-up, a stable archaeal community (similarity higher than 84 %) was observed and maintained throughout the continuous operation. This community exhibited high LCFA tolerance and high acetoclastic and hydrogenotrophic activity. Cloning and sequencing results showed that Methanobacterium- and Methanosaeta-like microorganisms prevailed in the system and were able to tolerate and endure during prolonged exposure to high LCFA loads, despite the previously reported LCFA sensitivity of methanogens.     

Microbial community in granules from a high-rate EGSB reactor

The spatial distribution, quantity and diversity of different microorganisms within anaerobic granular sludge from a lab-scale expanded granular sludge bed (EGSB) reactor operated at different organic loading rates were studied using florescent in situ hybridization (FISH), real time quantitative — polymerase chain reaction (RTQ-PCR) and denaturing gradient gel electrophoresis (DGGE) techniques. The results indicated that most Eubacteria were located in the outer layer of granule, while the Archaea which mainly were methanogens and more sensible to the environmental conditions were located in the inner layer of the granule. The quantity of Archaea was obviously less than that of Eubacteria in the granules, but increased with the increasing of organic loading rates of the reactor. As the organic loading rate of the reactor increased and the operating time elapsed, the Archaea community in the granules changed significantly. Seven typical DGGE bands were collected and sequenced, and found that the dominant species of Archaea in the granules operated in the last period were mainly Methanocorpusculum, Methanobacterium, Methanosaeta.     

Performance of upflow anaerobic sludge blanket (UASB) reactor treating wastewaters containing carbon tetrachloride

The anaerobic biodegradation of carbon tetrachloride (CT) was investigated during the granulation process by reducing the hydraulic retention time, increasing the chemical oxygen demand (COD) and CT loadings in a 2l laboratory-scale upflow anaerobic sludge blanket (UASB) reactor. Anaerobic unacclimated sludge and glucose were used as seed and primary substrate, respectively. Granules were developed 4 weeks after start-up, which grew at an accelerated rate for 8 months, and then became fully grown. The effect of operational parameters such as influent CT concentrations, COD, CT loading, food to biomass ratio and specific methanogenic activity (SMA) were also considered during granulation. The granular sludge cultivated had a maximum diameter of 2.1 mm and SMA of 1.6 g COD/g total suspended solid (TSS) day. COD and CT removal efficiencies of 92 and 88% were achieved when the reactor was firstly operating at CT and COD loading rates of 17.5 mg/l day and 12.5 g/l day, respectively. This corresponds to hydraulic retention time of 0.28 day and food to biomass ratio of 0.5 g COD/g TSS day. Kinetic coefficients of maximum specific substrate utilization rate, half velocity coefficient, growth yield coefficient and decay coefficient were determined to be 2.4 × 10^–3 mg CT/TSS day^–1, 1.37 mg CT/l, 0.69 mg TSS/mg CT and 0.046 day^–1, respectively for CT biotransformation during granulation.     

Treatment of low strength industrial cluster wastewater by anaerobic hybrid reactor

The study was aimed at treating the complex, combined wastewater generated in Mangolpuri industrial cluster. It was considered as a low strength wastewater with respect to its organic content. Anaerobic treatment of this wastewater was studied using an anaerobic hybrid reactor (AHR) which combined the best features of both the upflow anaerobic sludge blanket (UASB) reactor and anaerobic fluidized bed rector (AFBR). The performance of the reactor under different organic and hydraulic loading rates were studied. The COD removal reached 94% at an organic loading rate (OLR) of 2.08 kg COD m(-3)d(-1) at an hydraulic retention time (HRT) of 6.0 h. The granules developed were characterized in terms of their diameter and terminal settling velocity.     

Molecular analysis of the biomass of a fluidized bed reactor treating synthetic vinasse at anaerobic and micro-aerobic conditions

The microbial communities (Bacteria and Archaea) established in an anaerobic fluidized bed reactor used to treat synthetic vinasse (betaine, glucose, acetate, propionate, and butyrate) were characterized by denaturing gradient gel electrophoresis (DGGE) and phylogenetic analysis. This study was focused on the competitive and syntrophic interactions between the different microbial groups at varying influent substrate to sulfate ratios of 8, 4, and 2 and anaerobic or micro-aerobic conditions. Acetogens detected along the anaerobic phases at substrate to sulfate ratios of 8 and 4 seemed to be mainly involved in the fermentation of glucose and betaine, but they were substituted by other sugar or betaine degraders after oxygen application. Typical fatty acid degraders that grow in syntrophy with methanogens were not detected during the entire reactor run. Likely, sugar and betaine degraders outnumbered them in the DGGE analysis. The detected sulfate-reducing bacteria (SRB) belonged to the hydrogen-utilizing Desulfovibrio. The introduction of oxygen led to the formation of elemental sulfur (S⁰) and probably other sulfur compounds by sulfide-oxidizing bacteria (γ-Proteobacteria). It is likely that the sulfur intermediates produced from sulfide oxidation were used by SRB and other microorganisms as electron acceptors, as was supported by the detection of the sulfur respiring Wolinella succinogenes. Within the Archaea population, members of Methanomethylovorans and Methanosaeta were detected throughout the entire reactor operation. Hydrogenotrophic methanogens mainly belonging to the genus Methanobacterium were detected at the highest substrate to sulfate ratio but rapidly disappeared by increasing the sulfate concentration.     

Anaerobic treatment of synthetic medium-strength wastewater using a multistage biofilm reactor

A laboratory-scale multistage anaerobic biofilm reactor of three compartments with a working volume of 54-L was used for treating a synthetic medium-strength wastewater containing molasses as a carbon source at different influent conditions. The start-up period, stability and performance of this reactor were assessed at mesophilic temperature (35 °C). During the start-up period, pH fluctuations were observed because there was no microbial selection or zoning, but as the experiment progressed, results showed that phase separation had occurred inside the reactor. COD removal percentages of 91.6, 91.6, 90.0 and 88.3 were achieved at organic loading rates of 3.0, 4.5, 6.75 and 9.0 kg COD/m³ day, respectively. A decrease in HRT from 24 to 16 h had no effect on COD removal efficiency. When HRT decreased to 8 h, COD removal efficiency was still 84.9%. Recirculation ratios of 0.5 and 1.0 had no effect on COD removal but other factors such as the volatile fatty acid (VFA) content were affected. The effect of toxic shock was also investigated and results showed that the main advantage of using this bioreactor lies in its compartmentalized structure.     

Treatment of distillery spentwash by hybrid UASB reactor

A laboratory-scale hybrid UASB reactor, which combined an UASB in the lower part and a filter in the upper part, was used for the treatment of distillery spentwash. The reactor was operated under ambient conditions for 380 days. Using anaerobically digested sewage sludge as a seed, the start-up of the reactor and the cultivation of active granular sludge was completed within three months period. Scanning electron microscopic (SEM) observation of the granules showed the presence of Mehtanonthrix-like bacteria as the dominant species. Following the start-up the organic loading rate (OLR) was increased, stepwise, to 36 kg COD/m³ · d at a constant hydraulic retention time (HRT) of 6 h. COD removal efficiency was 80% even at a high OLR of 36 kg COD/m³ · d. Biogas rich in methane content (80%), with a maximum specific biogas yield of 0.40 m³ CH₄/kg · COD was produced. Polypropylene pall rings filter medium in the upper-third of the reactor was very effective as a gas-liquid-solid (GLS) separator, and retained the biomass in addition. The study indicated that hybrid UASB is a very feasible alternative for the treatment of high-strength wastewaters like distillery spentwash.     

Anaerobic digestion using a fixed-bed reactor with bacteria immobilized by polyacrylate on a saddle-shaped slag support

Anaerobic bacteria were immobilized by polyacrylate, which was cross-linked with Ca²⁺ ion, on a saddle-shaped slag support. The substance diffusion characteristics were improved because of a thin polymer gel layer only 0.95 mm in thickness. After immobilization, the support was packed in a fixed-bed reactor which was recycled, and the reactor performance was evaluated by using a synthetic wastewater of glucose and cornsteep-liquor. The COD removal reached 90% on the 17th d, and the acclimation of the immobilized bacteria was accomplished within 30 d. The COD removal at an HRT of 24 h was 96 to 93% for a COD load of 5 to 12 gdm⁻³d⁻¹, COD removal of 85% was obtained at an HRT of 8 h and COD loading rate of 7.2 gdm⁻³d⁻¹. The SS in the effluent was 0.07 to 0.37 gdm⁻³, and the net biomass yield and methane conversion ratio were 0.02 gVSSg⁻¹ COD and 0.36 dm³g⁻¹ COD (25°C), respectively. In terms of system stability at a short HRT, methane conversion, and biomass yield, the performance of the immobilized system was better than the biofilm system.     

Effect of nickel deprivation on methanol degradation in a methanogenic granular sludge bioreactor

The effect of omitting nickel from the influent on methanol conversion in an Upflow Anaerobic Sludge Bed (UASB) reactor was investigated. The UASB reactor (30°C, pH 7) was operated for 261 days at a 12-h hydraulic retention time (HRT) and at organic loading rates (OLRs) ranging from 2.6 to 7.8 g COD l reactor⁻¹ day⁻¹. The nickel content of the sludge decreased by 66% during the 261-day reactor run because of washout and doubling of the sludge bed volume. Nickel deprivation initially had a strong impact on the methanogenic activity of the sludge with methanol; e.g., after 89 days of operation, this activity was doubled by adding 2 μM nickel. Upon prolonged UASB reactor operation, methanol and VFA effluent concentrations decreased whereas the sludge lost its response to nickel addition in activity tests. This suggests that a less nickel-dependent methanol-converting sludge had developed in the UASB reactor. Received 09 April 2002/ Accepted in revised form 13 July 2002     

The influence of anaerobic pretreatment on the nitrogen removal from biosynthetic pharmaceutical wastewaters

The C:N ratio of the pharmaceutical wastewaters is usually suitable for a combination of the anaerobic pretreatment with the high COD removal and aerobic posttreatment with the efficient biological N removal. This kind of anaerobic-aerobic process was tested in semipilot scale by using a UASB reactor and an activated sludge system with a predenitrification (total volume 100 1). It was found that at a total HRT of 2.3 days an average of 97.5% of COD and 73.5% of total N was removed. The UASB reactor was operated at 30°C with a volumetric loading rate of 8.7 kg.m^-3.d^-1, the efficiency of COD removal was 92.2%. The processes, which take part in the biological removal of nitrogen, especially the nitrification, were running with lower rates than usually observed in aerobic treatment systems.Abbreviations AAO anaerobic anoxic oxic configuration - AOO anaerobic oxic oxic configuration - B _V volumetric organic loading rate (kg COD.m^-3. d^-1) - dB _x specific COD removal rate (mg COD. g^-1 VSS. d^-1) - DNR denitrification rate (mg N–NO₃. g^-1 VSS. h^-1) - E_COD efficiency of COD removal (%) - HRT hydraulic retention time (d) - NR nitrification rate (mg N–NO₃. g^-1 VSS. h^-1) - R recirculation ratio (%) - SBP specific biogas production (m³.kg^-1 removed COD) - SRT solids retention time; sludge age (d) - SS suspended solids (g.1^-1) - UASB upflow anaerobic sludge blanket reactor - VSS volatile suspended solids (g.1^-1)     

Mechanical shear contributes to granule formation resulting in quick start-up and stability of a hybrid anammox reactor

It appears that if suspended biomass washout can be reduced effectively, granule formation will be fastened in fluidized bed. Quicker reactor start-up can be anticipated especially for those system keeping slow growth bacteria such as anammox. A hybrid reactor combined fixed-bed with nonwoven fabrics as biomass carrier and fluidized bed with slow speed mechanical stirring was therefore developed, and its nitrogen removal performances was evaluated experimentally. Only in 38 days, the total nitrogen removal rate (NRR) reached to 1.9 kg(N) m⁻³ day⁻¹ and then doubled within 17 days, with total nitrogen removal efficiency kept above 70%. After 180 days reactor operating, the NRR reached a maximum value of 6.6 kg(N) m⁻³ day⁻¹ and the specific anammox activity was gradually constant in 0.32 kg(N) kg(VSS)⁻¹ day⁻¹. Biomass attached on nonwoven fabrics could additionally improve reactor nitrogen removal by 8%. The dominant size of granular sludge reached to 0.78 mm with stirring speed adjusted from 30 to 80 rpm and the hydraulic retention time (HRT) from 8 to 1.5 h during the whole operating time. Scanning electron microscope observation showed especially compact structure of granular sludge. A 70% of anammox bacteria percentage was identified by fluorescence in situ hybridization analysis.     

Comparison of startup and anaerobic wastewater treatment in UASB, hybrid and baffled reactor

An experimental study was carried out to compare the performance of selected anaerobic high rate reactors operated simultaneously at 37?°C. The three reactors, namely upflow anaerobic sludge bed reactor (UASB), hybrid of UASB reactor and anaerobic filter (anaerobic hybrid reactor – AHR) and anaerobic baffled reactor (ABR), were inoculated with the anaerobic digested sludge from municipal wastewater treatment plant and tested with synthetic wastewater. This wastewater contained sodium acetate and glucose with balanced nutrients and trace elements (COD 6000?mg?·?l^?1). Organic loading rate (B _v ) was increased gradually from an initial 0.5?kg?·?m^?3?·?d^?1 to 15?kg?·?m^?3?·?d^?1 in all the reactors. From the comparison of the reactors' performance, the lowest biomass wash-out resulted from ABR. In the UASB, significant biomass wash-out was observed at the B _v 6?kg?·?m^?3?·?d^?1, and in the AHR at the B _v 12?kg?·?m^?3?·?d^?1. The demand of sodium bicarbonate for pH maintenance in ABR was two times higher as for UASB and AHR. The efficiency of COD removal was comparable for all three reactors – 80–90%. A faster biomass granulation was observed in the ABR than in the other two reactors. This fact is explained by the kinetic selection of filamentous bacteria of the Methanotrix sp. under a high (over 1.5?g?·?l^?1) acetate concentration.