Single-stage anaerobic codigestion for mixture wastes of simulated Korean food waste and waste activated sludge

Korean food waste was treated with a single-stage anaerobic codigester (SSAD) using waste activated sludge (WAS) generated from a municipal wastewater treatment plant. The stability and performance of the system was analyzed. The C/N ratio was improved with increasing food waste fraction of feed mixture. The pH, alkalinity, and free ammonia nitrogen concentration were the parameters used to evaluate the digester’s stability. The experimentally determined values of the parameters indicated that there were no methane inhibitions in the digester. Digester performance was determined by measuring the total chemical oxygen demand TCOD), volate solids (VS) removal, methane content in biogas, methane production rate (MPR), and specific methane productivity. Methane content in biogas and MPR were significantly dependent on hydraulic retention time (HRT) and ratio of food waste to WAS. The methane content in biogas decreased at shorter HRT or higher organic loading rate (OLR) with increased food waste fraction. Concerning the performance of the codigester, the optimum operating condition of the SSAD was found to be at an HRT of 10 d with a feed mixture ratio of 50% food waste and 50% WAS. A TCOD removal efficiency of 53.6% and a VS removal efficiency of 53.7% were obtained at an OLR of 5.96 kg of TCOD/(m³·d) and 3.14 kg of VS/(m³·d), respectively. A maximum MPR of 1.15 m³ CH₄/(m³·d) and an SMP of 0.37 m³ CH₄/kg of VS_feed were obtained at an HRT of 10 d with a methane content of 63%.     

Anaerobic bioconversion of municipal solid wastes using a novel high-solids reactor design

Novel, laboratory-scale, high-solids reactors operated under mesophilic conditions were used to study the anaerobic fermentation of processed municipal solid waste (MSW) to methane. Product gas rate data were determined for organic loading rates ranging from 2.99–18.46 g of volatile solids (VS) per liter (L) per day (d). The data represent the anaerobic fermentation at high-solids levels within the reactor of 21–32%, while feeding a refuse-derived fuel (RDF)/MSW feedstock supplemented with a vitamin/mineral/nutrient solution. The average biogas yield was 0.59 L biogas/g VS added to the reactor system/d. The average methane composition of the biogas produced was 57.2%. The data indicate a linear relationship of increasing total biogas production with increasing organic loading rate to the process. The maximum organic loading rate obtainable with high-solids anaerobic digestion is in the range of 18–20 g VS/L·d to obtain 80% or greater bioconversion for the RDF/MSW feedstock. This loading rate is approximately four to six times greater than that which can be obtained with comparable low-solids anaerobic bioreactor technology.     

Evaluation of Biogas Production Potential by Dry Anaerobic Digestion of Switchgrass–Animal Manure Mixtures

Anaerobic digestion is a biological method used to convert organic wastes into a stable product for land application with reduced environmental impacts. The biogas produced can be used as an alternative renewable energy source. Dry anaerobic digestion [>15% total solid (TS)] has an advantage over wet digestion (<10% TS) because it allows for the use of a smaller volume of reactor and because it reduces wastewater production. In addition, it produces a fertilizer that is easier to transport. Performance of anaerobic digestion of animal manure–switchgrass mixture was evaluated under dry (15% TS) and thermophilic conditions (55 °C). Three different mixtures of animal manure (swine, poultry, and dairy) and switchgrass were digested using batch-operated 1-L reactors. The swine manure test units showed 52.9% volatile solids (VS) removal during the 62-day trial, while dairy and poultry manure test units showed 9.3% and 20.2%, respectively. Over the 62 day digestion, the swine manure test units yielded the highest amount of methane 0.337 L CH₄ /g VS, while the dairy and poultry manure test units showed very poor methane yield 0.028 L CH₄/g VS and 0.002 L CH₄/g VS, respectively. Although dairy and poultry manure performed poorly, they may still have high potential as biomass for dry anaerobic digestion if appropriate designs are developed to prevent significant volatile fatty acid (VFA) accumulation and pH drop.     

Enhancement of Biogas Production by Co-digestion of Potato Pulp with Cow Manure in a CSTR System

Anaerobic digestion (AD) process is a well-established method to generate energy from the organic wastes both from the environmental and economical perspectives. The purpose of present study is to evaluate energy production from potato wastes by incorporating cow manure into the process. Firstly, a laboratory pilot of one-stage biogas production was designed and built according to continuously stirred tank reactor (CSTR) system. The setup was able to automatically control the environmental conditions of the process including temperature, duration, and rate of stirring. AD experiment was exclusively performed on co-digestion of potato peel (PP) and cow manure (CM) in three levels of mixing ratio including 20:80, 50:50, 80:20 (PP:CM), and 0:100 as control treatment based on the volatile solid (VS) weight without adding initial inoculums. After hydraulic retention time (HRT) of 50 days on average 193, 256, 348, and 149 norm liter (L_N) (kg VS)^?1, methane was produced for different mixing ratios, respectively. Statistical analysis shows that these gas productions are significantly different. The average energy was determined based on the produced methane which was about 2.8 kWh (kg VS)^?1, implying a significant energy production potential. The average chemical oxygen demand (COD) removal of treatments was about 61 %, showing that it can be leached significantly with high organic matter by the employed pilot. The energy efficiency of 92 % of the process also showed the optimum control of the process by the pilot.     

Pretreatment of Chicken Feather Waste for Improved Biogas Production

This study deals with the utilization of chicken feather waste as a substrate for anaerobic digestion and improving biogas production by degradation of the compact structure of the feather keratin. In order to increase the digestibility of the feather, different pretreatments were investigated, including thermal pretreatment at 120 °C for 10 min, enzymatic hydrolysis with an alkaline endopeptidase [0.53–2.66 mL/g volatile solids (VS) feathers] for 0, 2, or 24 h at 55 °C, as well as a combination of these pretreatments. The effects of the treatments were then evaluated by anaerobic batch digestion assays at 55 °C. The enzymatic pretreatment increased the methane yield to 0.40 Nm³/kg VS_added, which is 122 % improvement compared to the yield of the untreated feathers. The other treatment conditions were less effective, increasing the methane yield by 11–50 %. The long-term effects of anaerobic digestion of feathers were examined by co-digestion of the feather with organic fraction of municipal solid waste performed with and without the addition of enzyme. When enzyme was added together with the feed, CH₄ yield of 0.485 Nm³/kg VS^?1 d^?1 was achieved together with a stable reactor performance, while in the control reactor, a decrease in methane production, together with accumulation of undegraded feather, was observed.     

Biogasification of Green and Food Wastes Using Anaerobic-Phased Solids Digester System

The performance of a laboratory-scale anaerobic-phased solid (APS) digester system treating food and green wastes was evaluated at thermophilic condition. The APS system comprised of four hydrolysis digesters and one biogasification reactor. The hydrolysis reactors were operated batchwised at a 12-day retention time, while the biogasification reactor was continuously operated at different hydraulic retention times (HRT). The biogas and methane yields from green waste were determined to be 0.438 and 0.252 L/g volatile solid (VS), respectively, with VS removal of 78%. The biogas and methane yields from food waste were 0.596 and 0.379 L/g VS, respectively, with VS removal of 85%. Hydrogen was produced from hydrolysis reactors during the digestion of food waste. Its content was 30.1% and 8.5% of the biogas produced on the first and second day of digestion, respectively. Hydrogen yield from the whole system was determined to be 0.029 L/g VS representing about 4.9% of the total biogas production from the system. The ratio between the volumes of biogasification and hydrolysis reactors (BR/HR) was found to be a factor that affects the process performance and stability.     

Two-phase methanization of food wastes in pilot scale

A 5 ton/d pilot scale two-phase anaerobic digester was constructed and tested to treat Korean food wastes in Anyang city near Seoul. The easily degradable presorted food waste was efficiently treated in the two-phase anaerobic digestion process. The waste contained in plastic bags was shredded and then screened for the removal of inert materials such as fabrics and plastics, and subsequently put into the two-stage reactors. Heavy and light inerts such as bones, shells, spoons, and plastic pieces were again removed by gravity differences. The residual organic component was effectively hydrolyzed and acidified in the first reactor with 5 d space time at pH of about 6.5. The second, methanization reactor converted the acids into methane with pH between 7.4 and 7.8. The space time for the second reactor was 15 d. The effluent from the second reactor was recycled to the first reactor to provide alkalinities. The process showed stable steady-state operation with the maximum organic loading rate of 7.9 kg volatile solid (VS)/m³/d and the volatile solid reduction efficiency of about 70%. The total of 3.6 tons presorted MSW containing 2.9 tons of food organic was treated to produce about 230 m³ of biogas with 70% (v/v) of methane and 80 kg of humus. This process is extended to full-scale treating 15 tons of food waste a day in Euiwang city and the produced biogas is utilized for the heating/cooling of adjacent buildings. Author to whom all correspondence and reprint requests should be addressed.     

Influence of Hydraulic Retention Time and Reactor Configuration During Fermentation of Diluted Chicken Manure

In this study, single-stage and two-phase semi-continuous thermophilic anaerobic reactors fed with diluted (3 % total solids (TS) and 1.8 % volatile solids (VS)) chicken manure at three different hydraulic retention times (HRTs) were compared interms of biogas production rate, methane content of the produced biogas, and VS and TS removal. Along the study, HRTs of 16, 12, and 8 days were implemented to the single-stage and the two-phase systems. It was observed that the single-stage anaerobic system was superior to the two-phase anaerobic system according to their biogas production rates (517 vs. 356, 551 vs. 359, 459 vs. 386 (mL/g VS_feed)) at all HRTs. On the other hand, methane content of the biogas produced was higher in the two-phase system compared to the single-stage system.     

Anaerobic treatment of animal byproducts from slaughterhouses at laboratory and pilot scale

Different mixtures of animal byproducts, other slaughterhouse waste (i.e., rumen, stomach and intestinal content), food waste, and liquid manure were codigested at mesophilic conditions (37°C) at laboratory and pilot scale. Animal byproducts, including blood, represent 70–80% of the total biogas potential from waste generated during slaughter of animals. The total biogas potential from waste generated during slaughter is about 1300 MJ/cattle and about 140 MI/pig. Fed-batch digestion of pasteurized (70°C, 1h) animal byproducts resulted in a fourfold increase in biogas yield (1.14L/g of volatile solids [VS]) compared with nonpasteurized animal bypproducts (0.31L/g of VS). Mixtures with animal byproducts representing 19–38% of the total dry matter were digested in continuous-flow stirred tank reactors at laboratory and pilot scale. Stable processes at organic loading rates (OLRs) exceeding 2.5g of VS/(L·d) and hydraulic retention times (HRTs) less than 40 d could be obtained with total ammonia nitrogen concentrations (NH₄−N+NH₃−N) in the range of 4.0–5.0 g/L. After operating one process for more than 1.5 yr at total ammonia nitrogen concentrations >4 g/L, an increase in OLR to 5 g of VS/(L·d) and a decrease in HRT to 22 d was possible without accumulation of volatile fatty acids.     

Process Improvement of Biogas Production from Anaerobic Co-digestion of Cow Dung and Corn Husk

A study of organic loading rate (OLR) and effect of NaOH concentration on the pretreatment stage of corn husk (CH) was conducted by Face-Centered Central Composite Design (CCF) to improve the biogas production. Three levels of OLR at 25.0 g VS · L⁻¹ · d⁻¹ (OLR₂₅), 35.0 g VS · L⁻¹ · d⁻¹ (OLR₃₅), and 45.0 g VS · L⁻¹ · d⁻¹ (OLR₄₅) were performed with NaOH pretreatment concentration of CH at 25.0% (N₂₅), 35.0% (N₃₅), and 45.0% (N₄₅) (w/w). The optimum production of biogas at 67.6 mL · min⁻¹ with methane concentration of 63.4% has been obtained at the application of OLR at 43.6 g VS · L⁻¹ · d⁻¹ and NaOH concentration at 33% (w/w).     

Dry Anaerobic Co-digestion of Cow Dung with Pig Manure for Methane Production

The performance of dry anaerobic digestions of cow dung, pig manure, and their mixtures into different ratios were evaluated at 35?±?1 °C in single-stage batch reactors for 63 days. The specific methane yields were 0.33, 0.37, 0.40, 0.38, 0.36, and 0.35 LCH₄/gVS_r for cow dung to pig manure ratios of 1:0, 4:1, 3:2, 2:3, 1:4, and 0:1, respectively, while volatile solid (VS) and chemical oxygen demand (COD) removal efficiencies were 48.59, 50.79, 53.20, 47.73, 46.10, and 44.88 % and 55.44, 57.96, 60.32, 56.96, 53.32, and 50.86 %, respectively. The experimental results demonstrated that the co-digestions resulted in 5.10–18.01 % higher methane yields, 2.03–12.95 % greater VS removals, 2.98–12.52 % greater COD degradation and so had positive synergism. The various mixtures of pig manure with cow dung might persuade a better nutrient balance and dilution of high ammonia concentration in pig manure and therefore enhanced digester performance efficiency and higher biogas yields. The dry co-digestion of 60 % cow dung and 40 % pig manure achieved the highest methane yield and the greatest organic materials removal efficiency than other mixtures and controls.     

Evaluation of the Anaerobic Co-Digestion of Sewage Sludge and Tomato Waste at Mesophilic Temperature

Sewage sludge is a hazardous waste, which must be managed adequately. Mesophilic anaerobic digestion is a widely employed treatment for sewage sludge involving several disadvantages such as low methane yield, poor biodegradability, and nutrient imbalance. Tomato waste was proposed as an easily biodegradable co-substrate to increase the viability of the process in a centralized system. The mixture proportion of sewage sludge and tomato waste evaluated was 95:5 (wet weight), respectively. The stability was maintained within correct parameters in an organic loading rate from 0.4 to 2.2 kg total volatile solids (VS)/m³ day. Moreover, the methane yield coefficient was 159 l/kg VS (0 °C, 1 atm), and the studied mixture showed a high anaerobic biodegradability of 95 % (in VS). Although the ammonia concentration increased until 1,864?±?23 mg/l, no inhibition phenomenon was determined in the stability variables, methane yield, or kinetics parameters studied.     

Optimization of inoculum to substrate ratio for enhanced methane yield from leather fleshings in a batch study

One of the most significant issues of the last few decades has been tracing for renewable energy sources. Animal fleshing (ANFL) is the most common proteinaceous solid waste accured during the production of leather and it must be disposed of in an environmentally responsible manner. This paper is attempts to assess the biogas production from solid waste originating from the chrome based tannery. Anaerobic digestion of these wastes will be a viable option for waste stabilization and energy production in the form of biogas to be utilized in the industry. The bio-methane potential of the wastes were examined by mixing these wastes with various sources of inoculum and different inoculum to substrate (I/S) ratio considered. The batch experiments were carried out in 2.5 l glass reactors with a various source of inoculumviz., Cow Dung (CD), Elephant Dung (ED) and Bio-Digested Slurry (BDS) with varied inoculum to substrate (LFs) ratios for a retention time of 50 days with replications. The results obtained from the experiments showed that BDS:LF (25:75) had the highest gas production of 14505 ml (651.85 ml CH₄g^?1 VS) followed by CD:LF (50:50) produced 12072.5 ml (789.36 ml CH₄g^?1 VS) and ED:LF (75:25) produced 11252.5 ml (1492.08 ml CH₄g^?1 VS)with a methane content of 63.77, 61.92 and 62.72%, respectively.     

Evaluating the biochemical potential for Irbid’s food waste as a source for energy

Solid waste generated from the city of Irbid is all directed to a local unlined landfill with no gas collection system. More than half of the waste stream is made of food waste which turns the landfill to a concentrated point source of greenhouse gases. To assess the impact that the landfill has at the moment on global warming and to evaluate the future value of the city’s waste as a source of biogas, 20 trucks of municipal solid waste were sampled. In addition, food waste samples from Irbid were collected from five different sources for 12 weeks to quantify their generation rates and to measure their components and chemical composition. Irbid’s solid waste was found to have a distinctively high biodegradable content 68%, which is typical of developing countries’ solid waste. Physical assessment showed that 38%, 6%, 33% and 23% of the food waste was rice, meat, fruits and vegetables, and bread, respectively. Measured methane yields for the volatile solids (VS) in rice, meat, fruits and vegetables, and bread were 362, 499, 352 and 375 mL/g VS, respectively. A representative food waste sample was created to test the actual methane yield and compare it to calculated one. Actual methane yield (414 mL/g VS) was greater than the calculated value (370 mL/g VS) based on food type proportions and their specific methane yield. To assess the anaerobic biodegradability of food waste, a stoichiometric calculation of the methane production was made based on the elemental analysis of all food waste elements. Food waste was found to have the chemical formula C₇H₁₆O₅N and calculated methane potential of 447 mL/g VS; indicating 85% anaerobic biodegradability of food waste, which makes Irbid’s waste perceived as a valuable source of energy.     

Inhibition mechanisms of ammonia and sulfate in high-solids anaerobic digesters for food waste treatment: Microbial community and element distributions responses

The horizontal flow anaerobic digester indicated that high ammonia (2923 mg/L) and SO₄^2? (3653 mg/L) would influence the performance of methane production with food waste as substrates. Therefore, bottle anaerobic digestion reactors were carried out to investigate the effect of ammonia/sulfate concentrations on the methane production. Experimental results manifested that the anaerobic digesters with an ammonia concentration of 3500 mg/L or sulfate of 1600 mg/L showed the best performance of methane production, with an average methane yield of 0.32 and 0.33 L (g VS)^?1 d^?1, respectively. Specifically, a higher ammonia (6500 mg/L) or sulfate (1600-3500 mg/L) level hindered the bioconversion of C from liquid to gas phase (2.68% or 1.73% CH₄-Gas, respectively), while insignificantly for the hydrolyzation of C and N from solid to liquid phase. Similar to sulfate, high ammonia nitrogen seriously inhibited the methanation process, leading to a significant carbon accumulation in the anaerobic reactor, especially for propionic acid. The predominant archaea Methanosarcina at genus level indicated that aceticlastic methanogenesis was the major methanogenic pathway. Meanwhile, high ammonia level suppressed the activity of Methanosarcina, while modest sulfate improved H₂-consuming methanogens activity. A large fraction of unclassified bacteria within the Firmicutes (43.78%-63.17%) and Bacteroidetes (24.20%-33.30%) phylum played an important role in substrates hydrolysis.     

Optimization of Anaerobic Co-digestion of Strawberry and Fish Waste

Anaerobic co-digestion of agri-food waste is a promising management alternative. Its implementation, however, requires evaluating the proportion in which waste should be mixed to optimize their centralized treatment. The combined treatment of strawberry extrudate and fish waste, which are widely generated in Mediterranean areas, was optimized. Strawberry extrudate and fish waste were mixed and treated at different proportions (88:12, 94:6, and 97:3, respectively; wet basis). The proportions selected for the mixture allow the different flows to be absorbed simultaneously. The highest methane production was observed for the ratio 94:6 (0.205 m³ _STP CH₄/kg volatile solid) (VS) (STP; 0 °C, 1 atm), with a methane production rate in the range of 5?·?10^?3–9?·?10^?3 m³ _STP/kg VS?·?d, while the highest organic loading rate was observed for the mixture at a proportion 88:12 (1.9?±?0.1 kg VS/m³?·?d). Biodegradability was found to be similar for the 88:12 and 94:6 proportions, with values around 90 % in VS. Nevertheless, the 97:3 ratio was not viable due to a low methane production. An inhibition phenomenon occurred at increasing loads due to the effect of some compounds contained in the fish waste such as chloride or nitrogen.     

Synergistic Effects of Magnetic Nanomaterials on Post-Digestate for Biogas Production

Digestate is characterized by high water content, and in the water and wastewater treatment settings, necessitates both large storage capacities and a high cost of disposal. By seeding digestate with four magnetic nanoparticles (MNPs), this study aimed to recover biogas and boost its methane potential anaerobically. This was carried out via biochemical methane potential (BMP) tests with five 1 L bioreactors, with a working volume of 80% and 20% head space. These were operated under anaerobic conditions at a temperature 40 °C for a 30 d incubation period. The SEM/EDX results revealed that the morphological surface area of the digestate with the MNPs increased as compared to its raw state. Comparatively, the degree of degradation of the bioreactors with MNPs resulted in over 75% decontamination (COD, color, and turbidity) as compared to the control system result of 60% without MNPs. The highest biogas production (400 mL/day) and methane yield (100% CH₄) was attained with 2 g of Fe₂O₄-TiO₂ MNPs as compared to the control biogas production (350 mL/day) and methane yield (65% CH₄). Economically, the highest energy balance achieved was estimated as 320.49 ZAR/kWh, or 22.89 USD/kWh in annual energy savings for this same system. These findings demonstrate that digestate seeded with MNPs has great potential to improve decontamination efficiency, biogas production and circular economy in wastewater management.     

Efficiency of Fe3O4 Nanoparticles with Different Pretreatments for Enhancing Biogas Yield of Macroalgae Ulva intestinalis Linnaeus

In this work, different pretreatment methods for algae proved to be very effective in improving cell wall dissociation for biogas production. In this study, the Ulva intestinalis Linnaeus (U. intestinalis) has been exposed to individual pretreatments of (ultrasonic, ozone, microwave, and green synthesized Fe₃O₄) and in a combination of the first three mentioned pretreatments methods with magnetite (Fe₃O₄) NPs, (ultrasonic-Fe₃O₄, ozone-Fe₃O₄ and microwave-Fe₃O₄) in different treatment times. Moreover, the green synthesized Fe₃O₄ NPs has been confirmed by FTIR, TEM, XRD, SEM, EDEX, PSA and BET. The maximum biogas production of 179 and 206 mL/g VS have been attained when U. intestinalis has been treated with ultrasonic only and when combined microwave with Fe₃O₄ respectively, where sediment were used as inoculum in all pretreatments. From the obtained results, green Fe₃O₄ NPs enhanced the microwave (MW) treatment to produce a higher biogas yield (206 mL/g VS) when compared with individual MW (84 mL/g VS). The modified Gompertz model (R² = 0.996 was appropriate model to match the calculated biogas production and could be used more practically to distinguish the kinetics of the anaerobic digestion (AD) period. The assessment of XRD, SEM and FTIR discovered the influence of different treatment techniques on the cell wall structure of U. intestinalis.     

Biological Pretreatment of Chicken Feather and Biogas Production from Total Broth

Chicken feathers are available in large quantities around the world causing environmental challenges. The feathers are composed of keratin that is a recalcitrant protein and is hard to degrade. In this work, chicken feathers were aerobically pretreated for 2–8 days at total solid concentrations of 5, 10, and 20 % by Bacillus sp. C₄, a bacterium that produces both α- and β-keratinases. Then, the liquid fraction (feather hydrolysate) as well as the total broth (liquid and solid fraction of pretreated feathers) was used as substrates for biogas production using anaerobic sludge or bacteria granules as inoculum. The biological pretreatment of feather waste was productive; about 75 % of feather was converted to soluble crude protein after 8 days of degradation at initial feather concentration of 5 %. Bacteria granules performed better during anaerobic digestion of untreated feathers, resulting in approximately two times more methane yield (i.e., 199 mlCH₄/gVS compared to 105 mlCH₄/gVS when sludge was used). Pretreatment improved methane yield by 292 and 105 % when sludge and granules were used on the hydrolysate. Bacteria granules worked effectively on the total broth, yielded 445 mlCH₄/gVS methane, which is 124 % more than that obtained with the same type of inoculum from untreated feather.     

Anaerobic baffled reactor treatment of biodiesel-processing wastewater with high strength of methanol and glycerol: reactor performance and biogas production

Biodiesel-processing factories employing the alkali-catalyzed transesterification process generate a large amount of wastewater containing high amount of methanol, glycerol, and oil. As such, wastewater has high potential to produce biogas using anaerobic treatment. The aim of this research was to investigate the performance of an anaerobic baffled reactor for organic removal and biogas production from biodiesel wastewater. The effect of different organic loading rates, varying from 0.5 kg m⁻³ d⁻¹ to 3.0 kg m⁻³ d⁻¹ of chemical oxygen demand, was determined using three 22 L reactors, each comprising five separate compartments. Wastewater was pretreated with chemical coagulants to partially remove oil prior to experimentation. Results show that the anaerobic baffled reactor operated at 1.5 kg m⁻³ d⁻¹ of chemical oxygen demand and ten days of hydraulic retention time provided the best removal efficiencies of 99 % of chemical oxygen demand, 100 % of methanol, and 100 % of glycerol. Increasing the organic loading rate over 1.5 kg m⁻³ d⁻¹ of chemical oxygen demand led to excessive accumulation of volatile fatty acids thereby making the pH drop to a value unfavorable for methanogenesis. The biogas production rate was 12 L d⁻¹ and the methane composition accounted for 64–74 %. Phase-separated characteristics revealed that the highest chemical oxygen demand removal percentage was achieved in the first compartment and the removal efficiency gradually decreased longitudinally. A scanning electron microscopic study indicated that the most predominant group of microorganisms residing on the external surface of the granular sludge was Methanosarcina.