Mesophilic Digestion Research Articles

Mesophilic versus thermophilic digestion of sludge in anaerobic membrane bioreactors

Mesophilic versus thermophilic digestion of sludge in anaerobic membrane bioreactors

Impact of biochar on anaerobic digestion process and microbiome composition; focusing on pyrolysis conditions for biochar formation

This study is a comprehensive investigation of the role of biochar in anaerobic digestion relating various physicochemical properties and the potential biodegradability of biochar to the digestion process. Seven biochars, produced from cedar wood, wheat straw, digestate, and municipal sewage sludge, at temperatures ranging from 400 to 950 °C, were evaluated during mesophilic digestion of cellulose as a model substrate. Pyrolysis conditions significantly affected biochar's properties, with high temperatures resulting in larger surface areas (up to 1262 m2/g) and pore sizes (up to 0.7 cm3/g), more organized graphene-like structures, and higher electrical conductivities. Irrespective of the concentration and diversity of their physicochemical properties, most tested biochars did not affect methane production, despite enriching the relative abundance of the methanogenic community, even up to 42.7 %. Interestingly, wood-based biochar, post-treated by gasification at a high temperature (800 °C), reduced methane yield by up to 52 %. Contrary, methane was increased by 40 % due to residual biodegradable organic matter in biochar produced at a low temperature (400 °C) under incomplete anoxic conditions. This indicates that once the stoichiometric methane potential is reached, no additional methane can be produced by adding biochar, unless it acts as a co-substrate.

Effects of nano-bubble water on anaerobic co-digestion of cabbage waste and cow manure under mesophilic and thermophilic conditions

ABSTRACT The impact of four nano-bubble water (NBW) additions on the hydrolysis rate, methane yield, and microbial community of co-digestion of cabbage waste (CW) and cow manure (CM) under mesophilic and thermophilic conditions were investigated. Adding NBW under mesophilic conditions promoted hydrolysis, and the highest soluble chemical oxygen demand of the mesophilic digesters with the addition of CO2-NBW increased by 15.86%. Methane yield in the mesophilic digesters with Air-NBW and CO2-NBW increased by 17.54% and 14.72%, respectively. Moreover, the addition of NBW further accelerated the methane yield rate under mesophilic conditions. Due to the influence of thermophilic temperature, the impact of NBW addition on hydrolysis, methane yield, and methane yield rate in the thermophilic digesters did not differ significantly from the control. The addition of Air-NBW and N2-NBW in the thermophilic digesters resulted in only marginal increases in methane yield, by 1.09% and 5.61%, respectively. NBW addition enhanced both the abundance and diversity of microbial communities in both mesophilic and thermophilic digesters. The addition of NBW represents a promising technological advancement for enhancing the efficiency of anaerobic co-digestion of CW and CM.

Optimizing anaerobic digestion: Benefits of mild temperature transition from thermophilic to mesophilic conditions

Anaerobic digestion (AD) plays a significant role in renewable energy recovery. Upgrading AD from thermophilic (50–57 °C) to mesophilic (30–38 °C) conditions to enhance process stability and reduce energy input remains challenging due to the high sensitivity of thermophilic microbiomes to temperature fluctuations. Here we compare the effects of two decreasing-temperature modes from 55 to 35 °C on cell viability, microbial dynamics, and interspecies interactions. A sharp transition (ST) is a one-step transition by 20 °C d−1, while a mild transition (MT) is a stepwise transition by 1 °C d−1. We find a greater decrease in methane production with ST (88.8%) compared to MT (38.9%) during the transition period. ST mode overproduced reactive oxygen species by 1.6-fold, increased membrane permeability by 2.2-fold, and downregulated microbial energy metabolism by 25.1%, leading to increased apoptosis of anaerobes by 1.9-fold and release of intracellular substances by 2.9-fold, further constraining methanogenesis. The higher (1.6 vs. 1.1 copies per gyrA) metabolic activity of acetate-dependent methanogenesis implied more efficient methane production in a steady mesophilic, MT-mediated system. Metagenomic binning and network analyses indicated that ST induced dysbiosis in keystone species and greatly enhanced microbial functional redundancy, causing loss of microbial syntrophic interactions and redundant metabolic pathways. In contrast, the greater microbial interconnections (average degrees 44.9 vs. 22.1) in MT at a steady mesophilic state suggested that MT could better maintain necessary system functionality and stability through microbial syntrophy or specialized pathways. Adopting MT to transform thermophilic digesters into mesophilic digesters is feasible and could potentially enhance the further optimization and broader application of practical anaerobic engineering.

Analyzing the Effect of Combined Chemical Conditioning and pH Adjustment on Improving Dewatering and Phosphorus Recovery from Anaerobic Mesophilic Digestate

Analyzing the Effect of Combined Chemical Conditioning and pH Adjustment on Improving Dewatering and Phosphorus Recovery from Anaerobic Mesophilic Digestate

Ammonia stripping by in situ biogas self-circulation to upgrade continuous thermophilic and mesophilic digestion of hydrothermal high-solid sludge

High-solid anaerobic digestion of hydrothermal sewage sludge has been developed. In order to upgrade the process by focusing on ammonia inhibition, a simply-equipped stripping system without additional alkali or heat supply was introduced by in situ biogas self-circulation. As the determined limit of total ammonia nitrogen at 1500 mg/L and 1000 mg/L for the mesophilic (MAD) and thermophilic anaerobic digestion (TAD) respectively and stripping rate at 5 L/min, continuous MAD and TAD was conducted in parallel. The stripping system successfully polished up the ammonia inhibition, and methanogenic capability of the TAD was promoted to approximately 90.0 % of the potential. Intermittent stripping mode proved usable. More frequent stripping was inevitable for the TAD as compared to the MAD. Hydraulic retention time below 20 d resulted in failure of the stripping mode due to rapid ammonia generation. Overall, this technology was practical in upgrading high-solid sludge digestion by effective ammonia control.

Synergistic effect of hydrothermal sludge and food waste in the anaerobic co-digestion process: microbial shift and dewaterability.

While thermal hydrolysis technology is commonly employed for sewage sludge treatment in extensive wastewater treatment facilities, persistent challenges remain, including issues such as ammonia-induced digestive inhibition and reduced productivity stemming from nutrient deficiency within the hydrothermal sludge. In this study, the effects of hydrothermal sludge-to-food waste mixing ratios and fermentation temperatures on anaerobic co-digestion were systematically investigated through a semi-continuous experiment lasting approximately 100days. The results indicated that anaerobic co-digestion of hydrothermal sludge and food waste proceeded synergistically at any mixing ratio, and the synergistic effect is mainly attributed to the improvement of carbohydrate removal and digestive system stability. However, thermophilic digestion did not improve the anaerobic performance and methane yield. On the contrary, mesophilic digestion performed better in terms of organic matter removal, especially in the utilization of soluble carbohydrates, soluble proteins, and VFAs. Microbial community analysis revealed that the transition from mesophilic to thermophilic anaerobic co-digestion prompts changes in the methane-producing pathways. Specifically, the transition entails a gradual shift from pathways involving acetoclastic and hydrogenotrophic methanogenesis to a singular hydrogenotrophic methanogenesis pathway. This shift is driven by thermodynamic tendencies, as reflected in Gibbs free energy, as well as environmental factors like ammonia nitrogen and volatile fatty acids. Lastly, it is worth noting that the introduction of food waste did lead to a reduction in cake solids following dewatering. Nevertheless, it was observed that thermophilic digestion had a positive impact on dewatering performance.

Mesophilic, thermophilic and thermal hydrolysis process coupled anaerobic digestion of sewage sludge: Biomethane potential, pathogen removal and energy feasibility

Conventional Anaerobic digestion is one of the most preferred sludge management techniques however, it has limitations such as low hydrolysis rate, longer retention time, larger digester volume, and reduced methane generation etc., To overcome these problems, Thermal hydrolysis process (THP) has been used as an effective pretreatment method for enhancing methane generation. However, the effect of the thermal hydrolysis process in heavy metal solubilization and methane production from high SRT (solids retention time) sludge is still not clear. Anaerobic digestion of sewage sludge sourced from sequencing batch reactor (SBR) and conventional activated sludge process (CASP) systems were evaluated under mesophilic (MAD), thermophilic (TAD), and thermal hydrolysis process (THP) mediated anaerobic digestion (AD) in terms of methane generation and, VS and pathogen removal. THP resulted in almost 38% COD solubilization for SBR and CASP sludge. Anaerobic digestion of SBR sludge resulted in methane generation of 101, 166, and 390 mL/gVS under mesophilic, thermophilic, and THP pretreatment conditions, respectively. For CASP sludge, the highest methane generation was observed for THP pretreated sludge (587 mL/gVS), followed by thermophilic (298 mL/gVS) and mesophilic digestion (241 mL/gVS). The fecal coliforms in the mesophilic and thermophilic digested SBR sludge were 3000 and 620 MPN/g, respectively, while 6100 and 740 MPN/g were found in digested CASP sludge, respectively. The THP-MAD and TAD processed SBR, and CASP sludges met the Class A sludge fecal coliforms and Salmonella density criteria of <1000 MPN/g TS (dry basis) and 3 MPN/4g TS (dry basis), respectively, over MAD processed sludges. Even though THP pretreatment resulted in the release of heavy metals, the concentration of heavy metals in the THP effluent was lesser than the limits prescribed by USEPA. The energy demand of the THP unit (160 °C, 6 bar) was estimated to be 162 MJ/m3. The energy balance analysis revealed that THP pretreatment would lead to energy self-sufficiency upon integration in a wastewater treatment plant (WWTP). This study can provide an experimental basis to apply THP pretreatment to enhance hydrolysis rate in anaerobic digesters while maximizing methane production and achieving energy benefits over MAD and TAD.

COMPARING PHOSPHORUS RECOVERY AND OPTIMUM TIME DIGESTION FROM SINGLE AND MIXING SUBSTRATES BY USING ANAEROBIC MESOPHILIC DIGESTION TECHNIQUE

Phosphorus (P) is considered a limited and non-renewable mineral source that is essential in our daily lives, especially for the agricultural industry to produce fertilizer. Food Waste (FW), Palm Oil Mill Effluent Sludge (POMES), and Water Treatment Sludge (WTS) are wastes that are claimed to contain nutrients such as P. One of the methods that can be used to recover P is through the Anaerobic Mesophilic Digestion (AMD) technique. Therefore, this study was carried out to investigate the highest P recovery and time digestion from single and mixing substrates. The physicochemical and biological properties of raw FW, POMES, and WTS were determined, and it was reported that wastes were suitable to undergo the AMD process. Mesophilic conditions 350C (±1) were used as the appropriate conditions for the anaerobic bacteria (AB) to reproduce and grow. Results for single substrates found that POMES recovered the highest P with 57.88% (±1.24), and optimum digestion was at day 12. While FW and WTS recovered P with 31.94% (±0.76) and 26.47% (±1.51), respectively, the optimum digestion times for both wastes were at days 18 (FW) and 21 (WTS), respectively, which were longer than POMES. In mixing substrates, FW-POMES contributed to the highest P recovery value with 68.34% (±1.49), and the optimum time digestion was on day 9. Followed by WTS-POME, FW-WTS, and FW-POMES-WTS with values of 60.42% (±2.12) (day 15), 48.05% (±2.51) (day 8), and 20.85% (±1.33) (day 3), respectively. The better synergetic effect within anaerobic bacteria (AB) in mixing substrates with FW-POMES could eventually influence the highest P removal from the substrate in a short period of time. Moreover, this study also provides information on new strategies used in recovering P. Keyword(s) : Phosphorus recovery, Food Waste; Palm Oil Mill Effluent Sludge; Water Treatment Sludge; Anaerobic Mesophilic Digestion

Determination of operational parameters for the first stage of continuous temperature-phased anaerobic digestion of oily food waste: Influent concentration, hydraulic retention time, pH control and temperature

Anaerobic digestion is promising in both food waste reduction and energy upgrading. However, the intermediates in anaerobic digestion of such an oily food waste, long chain fatty acids, not only have low bioavailability but also significantly threaten operation stability. Temperature-phased anaerobic digestion is an emerging technology in recent years, expected to overcome those issues. Given significance of the first stage of the two-stage process, continuous thermophilic digestion of oily food waste was conducted, with mesophilic digestion operated in parallel as control, to determine practical operational parameters from operation performance and stability. The feeding concentration at approximately 4.50 % of volatile solids was identified to be applicative for a relatively stable operation. Due to slow β-oxidation for long chain fatty acids, hydraulic retention time for the first stage needed longer than 10 d to maintain anaerobes’ survival so that they were not inhibited. In addition to the risk from inhibition, readily acidified risk still existed, and instant pH control was demonstrated to be efficient in maintaining the habitat of microbial metabolism. Compared with no viable bacteria detected under uncontrolled pH, 36.7 % live cells could still be detected under controlled pH. Overall, thermophilic digestion obviously achieved advantages in higher substance degradation and methane production with lower stability risk. The genera Methanosarcina and Methanothermobacter played a significant role, with the ratio at about 2:1. While bacterial viability was not reduced remarkably in preliminary period of inhibition, long-term operation under high lipid loading resulted in its loss.

Use of a biogas-specific e-nose with machine learning to identify biogas pattern changes linked to hydraulic retention times in an anaerobic digester: A case study

Anaerobic digestion via biogas digesters is the most environmentally friendly solution to recycling degradable organic waste into organic fertilizer and energy. The organic fraction of municipal solid waste (OFMSW) is one of the main organic waste sources for biogas plants. The variations in characteristics of OFMSW as a main challenge lead to fluctuating organic loading rates and hydraulic retention time (HRT), which can adversely affect the performance of biogas plants. This study focused on the gaseous headspace of a biogas digester to assess a reliable monitoring method. An e-nose was applied to extract biogas patterns from a mesophilic digester, which operated at HRT of 30, 25, 20, and 15 days to treat OFMSW. The biogas samples were significantly classified based on their HRTs by the support vectors machine (SVM) and the random forest (RF). Finally, the results presented a promising method for monitoring the performance of the biogas digester.

Comparison of Thermophilic-Mesophilic and Mesophilic-Thermophilic Two-Phase High-Solid Sludge Anaerobic Digestion at Different Inoculation Proportions: Digestion Performance and Microbial Diversity.

This study investigated the performance of thermophilic-mesophilic (T-M) and mesophilic-thermophilic (M-T) two-phase sludge anaerobic digestion at different inoculation proportions after a change in digestion temperature. After temperature change, the pH, total ammonia nitrogen (TAN), free ammonia nitrogen (FAN), solubility chemical oxygen demand (SCOD), and total alkalinity (TA) levels of two-phase digesters were between thermophilic control digesters and mesophilic control digesters. However, the volatile fatty acid (VFA) levels of two-phase digesters were higher than those of thermophilic or mesophilic control digesters. The bacteria communities of M-T two-phase digesters were more diverse than those of T-M. After a change in digestion temperature, the bacterial community was dominated by Coprothermobacter. After a change of digestion temperature, the relative abundance (RA) of Methanobacterium, Methanosaeta, and Methanospirillum of M-T two-phase digesters was higher than that of T-M two-phase digesters. In comparison, the RA of Methanosarcina of T-M two-phase digesters was higher than that of M-T two-phase digesters. The ultimate methane yields of thermophilic control digesters were greater than those of mesophilic control digesters. Nevertheless, the ultimate methane yield levels of M-T two-phase digesters were greater than those of T-M two-phase digesters. The ultimate methane yields of all two-phase digesters presented an earlier increase and later decrease trend with the increasing inoculation proportion. Optimal methane production condition was achieved when 15% of sludge (T-M15) was inoculated under mesophilic-thermophilic conditions, which promoted 123.6% (based on mesophilic control) or 27.4% (based on thermophilic control). An optimal inoculation proportion (about 15%) balanced the number and activity of methanogens of high-solid sludge anaerobic digestion.

Iron and Hydrogen Peroxidation-Induced Post-Treatment Improvement of Municipal Mesophilic Digestate in an Alkaline Environment and Its Impact on Biosolids Quality

Challenges associated with mesophilic digestate (MD) involve volume, odor, and pathogens, which effective post-digestion treatments can address. The efficiency of MD post-treatment can be enhanced by conditioning with ferric chloride (FeCl3), hydrogen peroxide (H2O2), and polymer. This study aimed to observe the effect of combined chemical conditioning on volume reduction, phosphorus (P) release, odor, and pathogen reduction potential for MD. MD was conditioned with polymer only, polymer and FeCl3 at pH adjusted to 8.0 with lime (Ca(OH)2), and a blend of polymer, FeCl3, and hydrogen peroxide (H2O2) at pH 8.0. The results show that adding all three chemicals improved post-treatment efficiency at 2.1 kg/t DS FeCl3, 2.1 kg/t DS polymer, and 600 mg/L H2O2 at pH 8.0, compared with polymer or dual conditioning. At the combined dose, cake solid content, centrate P removal, and odor reduction capability improved compared with raw MD by 20%, 99%, and 66%, respectively. Combined chemical treatment reduced fecal coliform by 98% but does not fulfil class A requirements and showed 50% regrowth potential. The synergic effect of polymer, FeCl3, H2O2, and alkaline pH breakdown EPS, reduced water holding capacity and formed compacted flocs for better water removal and settling. This combination also precipitated P through FeCl3 while H2O2 oxidation curbs odor, enhancing further P removal from centrate.

Restructuring anaerobic hydrolysis kinetics in plant-wide models for accurate prediction of biogas production

In this study, the effect of anaerobic hydrolysis rate on biogas production was investigated with mesophilic digesters in seven large-scale wastewater treatment plants. A linear correlation was determined between the percentage of primary sludge mass in the total sludge fed to the digester and the overall anaerobic hydrolysis rate. The anaerobic hydrolysis rate of primary sludge was determined to be three times higher than that of biological sludge. The reduction factors for anaerobic hydrolysis (ηHYD,ana) were identified in the range of 0.11–0.30 which is lower compared to the recommended range (0.30–0.50) given in the literature. This study proposes a new model approach where anaerobic degradation kinetics of influent originated (XB) and decay originated (XB,E) particulate biodegradable organics are separated. Current plant-wide models with a single kinetic expression required recalibration of the model for calculating biogas flowrate for each treatment facility with different primary and secondary sludge ratios fed to the digesters. The new model structure is able to predict biogas production of all wastewater treatment plants without any recalibration effort by segregating degradation kinetics of two particulate biodegradable organic fractions (XB, XB,E).

Metagenomic approach reveals the mechanism of calcium oxide improving kitchen waste dry anaerobic digestion

In light of the characteristics of excessive acidification and low biogas yield during kitchen waste (KW) dry digestion, the impact of the calcium oxide (CaO) on KW mesophilic dry digestion was investigated, and the enhanced mechanism was revealed through metagenomic approach. The results showed that CaO increased the biogas production, when the CaO dosage was 0.07 g/g (based on total solid), the biogas production reached 656.84 mL/g suspended solids (VS), approximately 8.38 times of that in the control. CaO promoted the leaching and hydrolysis of key organic matter in KW. CaO effectively promoted the conversion of volatile fatty acid (VFA) and mitigated over-acidification. Macrogenome analysis revealed that CaO increased the microbial diversity in KW dry digestion and upregulated the abundance of genes related to amino acid and carbohydrates metabolism. This study provides an effective strategy with potential economic benefits to improve the bioconversion efficiency of organic matter in KW.

Achieving energy self-sufficiency in wastewater treatment plants by integrating municipal solid waste treatment: A process design study in Spain

Water-energy nexus is a highlighted topic nowadays, particularly, the energy consumption in wastewater treatment plants (WWTPs) is becoming an important issue. WWTPs typically consume more energy than the one that can be obtained from the biogas produced from sludge anaerobic digestion. In this work, a process-level analysis is presented to study the feasibility of integrating wastewater and municipal solid waste (MSW) treatment to achieve the energetically self-sustainable operation of a WWTP. The influence of the climate of different regions across the Iberian Peninsula on the energy requirements has also been evaluated. Mesophilic and thermophilic digestion are compared in Salamanca as a case base, and the optimal digestion temperature is also evaluated, finding a value of 30∘C. Moreover, in all cities considered, it is necessary for MSW to provide between 37% and 40% of the facility energy consumption corresponding to around 0.19 kg of MSW per kg of sludge, with a small difference between cities. From an economic point of view, an investment between 0.09 and 0.16 €/kg of sewage sludge is required for the integrated process. Therefore, this techno-economic assessment demonstrates the feasibility of integrating these two treatments for a fully self-sufficient and sustainable process.

Quantification of indicator and pathogenic bacteria in manures and digestates from three agricultural biogas plants over a one-year period

Interest in the conversion of manure in biogas via anaerobic digestion (AD) is growing, but questions remain about the biosafety of digestates. For a period of one year, we monitored the impact of three mesophilic agricultural biogas plants (BPs) mainly fed with pig manure (BP1, BP3) or bovine manure (BP2) on the physicochemical parameters, the composition of the microbial community and the concentration of bacteria (E. coli, enterococci, Salmonella, Campylobacter, Listeria monocytogenes, Clostridium perfringens, Clostridium botulinum and Clostridioides difficile). The BP2 digestate differed from those of the two other BPs with a higher nitrogen content, more total solids and greater abundance of Clostridia MBA03 and Disgonomonadacea. Persistence during digestion ranked from least to most, was: Campylobacter (1.6 to >2.9 log10 reduction, according to the BP) < E. coli (1.8 to 2.2 log10) < Salmonella (1.1 to 1.4 log10) < enterococci (0.2 to 1.2 log10) and C. perfringens (0.2 to 1 log10) < L. monocytogenes (-1.2 to 1.6 log10) < C. difficile and C. botulinum (≤0.5 log10). No statistical link was found between the reduction in the concentration of the targeted bacteria and the physicochemical and operational parameters likely to have an effect (NH3, volatile fatty acids and total solids contents, hydraulic retention time, presence of co-substrates), underlining the fact that the fate of the bacteria during mesophilic digestion depends on many interacting factors. The reduction in concentrations varied significantly over the sampling period, underlining the need for longitudinal studies to estimate the impact of AD on pathogenic microorganisms.

Effect of organic loading on anaerobic digestion of cow dung: Methane production and kinetic study

Organic loading influences the effectiveness of producing biogas through anaerobic digestion. This study set out to investigate the effect of organic loading on the anaerobic mesophilic digestion of cow dung, the parameters involved in the digestion process and to evaluate the kinetics. Anaerobic digestion of cow dung at different organic loading (gVS/L) of 14 gVS/L, 18gVS/L, 22 gVS/L, 26 gVS/L and 30 gVS/L were investigated. Increasing the organic loading increased the methane yield of the cow dung. The highest cumulative methane yield was observed at 30 gVS/L with 63.42 mL CH4/gVS while the highest biogas yield was reported at 192.53 mL/gVS with the highest methane content of 89%. In addition, the modified Gompertz model equation with an R2 of 0.9980 demonstrated strong consistency and a good fit between predicted and experimental data. The high number of substrates added to the systems when increasing the organic loading increased the λ and slow down the nutrient transport and hydrolysis. This study provides current information on the effects of organic loading on the anaerobic digestion of cow dung in batch mode, including experimental conditions and operational parameters.

Enhanced biomethane generation from the anaerobic digestion of wilted corn straw via control in mesophilic and thermophilic temperature intervals

Anaerobic digestion (AD) at mesophilic and thermophilic interval temperatures has received increasing attention due to an unexpectedly higher methane yield. However, there is no general consensus and the mechanisms of microbial regulation under interval temperature remain unclear. Therefore, this study evaluated the effect of conventional and interval temperatures on methanogenesis performance and microbial regulation in continuous AD of wilted corn straw. The results demonstrated that the hydrolysis rate increased with increasing temperatures, probably due to the predominant Firmicutes and Proteobacteria, whose abundance in D55 was more than 72% and 19%, respectively. However, in the stable period (31–55 d) experiments, digesters D35, D45, and D55 achieved an average methane yield of 224 ± 34, 311 ± 55, and 261 ± 49 N mL/g VS/d, respectively. Interval temperature substantially enhanced methane yield by 39.4% and 19.0% compared to that from mesophilic and thermophilic digesters, respectively. A higher proportion of Methanosarcina (94.42–94.73%) promoted the acetoclastic methanogenesis at interval temperature, implying that methanogenesis, rather than hydrolysis, is limited at the thermophilic temperatures because of the loss of Methanosarcina. Furthermore, a high abundance of essential genes involved in methane metabolism was found to stimulate methane production in the interval temperature digester. These findings revealed that the interval temperatures may be a balanced zone between hydrolysis and methanogenesis for achieving higher methane yield and provided a new perspective strategy for AD.

Application of a Full-Scale Horizontal Anaerobic Digester for the Co-Digestion of Pig Manure, Food Waste, Excretion, and Thickened Sewage Sludge

Many laboratory- and pilot-scale studies on anaerobic co-digestion have been conducted in Republic of Korea; however, studies on full-scale demonstration facilities are lacking. This study aimed to present a successful case of a large-scale anaerobic co-digestion facility in Republic of Korea for biogas generation from four organic wastes (pig manure, food waste, excretion, and thickened sewage sludge) using a horizontal anaerobic digester. A preliminary biochemical methane potential test was performed for the individual and mixed organic waste to design a treatment facility for 320 m3/day of organic waste generated in Seosan City. Subsequently, a horizontal anaerobic digester with a 35 day-retention time (based on 320 m3/day input) was constructed. Each organic waste was placed in an anaerobic reactor after pretreatment. The input was gradually increased after the first seeding, and the operation continued for 158 days. Total and volatile solids made up 4.1% and 3.3%, respectively. Throughout the operating period, the digester temperature was maintained at 35–40 °C for mesophilic digestion, and the pH was maintained at 7–8. The average organic matter removal efficiency (volatile solids basis) was 64% and the methane gas production rates were 0.35, 0.6, 0.26, 0.28, and 0.39 Nm3CH4/kg vs. for pig manure, food waste, excretion, thickened sewage sludge, and mixed waste, respectively, resulting in an average methane content of the biogas 68.8%.