Utilization strategy for algal bloom waste through co-digestion with kitchen waste: Comprehensive kinetic and metagenomic analysis.

Anaerobic co-digestion (AcoD) with kitchen waste (KW) is an alternative utilization strategy for algal bloom waste (AW). However, the kinetic characteristic and metabolic pathway during this process need to be explored further. This study conducted a comprehensive kinetic and metagenomic analysis for AcoD of AW and KW. A maximum co-digestion performance index (CPI) of 1.13 was achieved under the 12% AW addition. Co-digestion improved the total volatile fatty acids generation and the organic matter transformation efficiency. Kinetic analysis showed that the Superimposed model fit optimally (R2Adj = 0.9988-0.9995). The improvement of the kinetic process by co-digestion was mainly reflected in the increase of the methane production from slowly biodegradable components. Co-digestion enriched the cellulolytic bacterium Clostridium and the hydrogenotrophic methanogenic archaea Methanobacterium. Furthermore, for metagenome analysis, the abundance of key genes concerned in cellulose and lipid hydrolysis, pyruvate and methane metabolism were both increased in co-digestion process. This study provided a feasible process for the utilization of AW produced seasonally and a deeper understanding of the AcoD synergistic mechanism from kinetic and metagenomic perspectives.

Agronomic and phytotoxicity test with biosolids from anaerobic CO-DIGESTION with temperature and micro-organism phase separation, based on sewage sludge, vinasse and poultry manure.

This study deals with energy and agronomic valorisation by anaerobic co-digestion with temperature and microorganism phase separation of sewage sludge, vinasse and poultry manure, with the aim of achieving an integral waste management, obtaining bioenergy and biofertilizer that returns nutrients to the soil in a natural way. The yields obtained were 40 mL H2/gVS and 391 mLCH4/gVS. The resulting effluent showed more than 98 % removal of E. coli and Total Coliforms, as well as total removal of Salmonella. The results obtained in the phytotoxicity tests showed that all the proportions studied had phytostimulant and phytonutrient properties, with 20 % having the highest germination index (GI) with mean values of 145.30 %. Finally, the agronomic trial carried out with strawberry crops (Fragaria sp.) showed that the addition of this biosolid has fertilising properties and can be used as an agronomic amendment, with an increase of 145 % in fresh weight and 102.5 % in dry weight, and fruit production doubled with respect to the control. The ANOVA statistical study corroborated that there were significant differences in crop growth when applying different proportions of biofertilizer in the fertilizer. Therefore, these results show that this technology is promising and would contribute environmentally, socially and economically to the transfer towards a circular economy model.

Enhanced methane production from source-separated human feces (brown water) by single phase anaerobic co-digestion: Effects of different co-substrates.

Based on the concept of source separation of brown water (BW, human feces with flushing water) and yellow water (urine) in rural area, anaerobic co-digestion of BW with agricultural waste is a promising and effective method for rural waste treatment and resource recovery. The purpose of this study was to investigate the performance of different agricultural wastes (peanut straw (PST), peanut shell (PSH), swine wastewater acting as co-substrate for anaerobic co-digestion with BW, and the relative mechanisms were explored. When the mixed ratio was uniformly set as 1:1 (mass ratio, measured by volatile solid (VS)) and initial VS load as 20 g/L, the maximum cumulative methane production obtained by co-digestion (21 days) of BW and PST was 688 mL/g-VS, which performed better than the individual substrates (341 mL/g-VS), as well as the average of the sole BW and sole PST groups (531.2 mL/g-VS). The most impactful advantage was ascribed to the promotion of hydrolytic and acidogenic enzyme activities. The addition of PST also reduced the production of endogenous humus, which is difficult for biodegradation. Microbial community analysis showed that different co-substrates would affect the microbial community composition in the reactor. The relative abundance of hydrolytic acidogens in the PST and PSH co-digestion groups were higher than that in the SW co-digestion and sole BW groups, and the methanogenic archaea were dominated by the acetate-trophic Methanotrichaceae. The overall results suggest that anaerobic co-digestion is a feasible method, and co-digestion of BW and PST can improve methane production potential.

The dominant-substrate driven the enhanced performance in co-digestion of Pennisetum hybrid and livestock waste.

Co-digestion has been considered a promising method to improve methane yield. The effect of the proportion of dominant substrate on the performance and microbial community of anaerobic digestion of Pennisetum hybrid (PH) and livestock waste (LW) was investigated. An obvious synergistic effect was obtained with an increase of 15.20%-17.45% in specific methane yield compared to the predicted value. Meanwhile, the dominant substrate influenced the relational model between methane yield enhancement rate and mixture ratio. For the LW-dominant systems, a parabolic model between enhancement rate and mixture ratio was observed with a highest value of 392.16 mL/g VS achieved at a PH:LW ratio of 2:8. While a linear pattern appeared for PH-dominant systems with the highest methane yield of 307.59 mL/g VS. Co-digestion selectively enriched the relative abundance of Clostridium_sensu_stricto_1, Terrisporobacter, Syntrophomonas, Methanosarcina and Methanobacterium, which boosted the performance of hydrolysis, acidogenesis, acetogenesis and methanogenesis processes.

Microbial community changes and metabolic pathways analysis during waste activated sludge and meat processing waste anaerobic co-digestion.

Waste activated sludge (WAS) and meat processing waste (MPW) were acted as co-substrates in anaerobic co-digestion (AcD), and biochemical methane potential (BMP) test was carried out to investigate the methane production performances. Microbial community structure and metabolic pathways analyses were conducted by 16S rRNA high-throughput sequencing and functional prediction analysis. BMP test results indicated that AcD of 70% WAS+30% MPW and 50% WAS+50% MPW (VS/VS) could significantly improve methane yield to 371.05 mL/g VS and 599.61 mL/g VS, respectively, compared with WAS acting as sole substrate (191.87 mL/g VS). The results of microbial community analysis showed that Syntrophomonas and Petrimonas became the dominant bacteria genera, and Methanomassiliicoccus and Methanobacterium became the dominant archaea genera after MPW addition. 16S functional prediction analysis results indicated that genes expression of key enzymes involved in syntrophic acetate oxidation (SAO), hydrogenotrophic and methylotrophic methanogenesis were up-regulated, and acetoclastic methanogenesis was inhibited after MPW addition. Based on these analyses, it could be inferred that SAO combined with hydrogenotrophic and methylotrophic methanogenesis was the dominant pathway for organics degradation and methane production during AcD. These findings provided systematic insights into the microbial community changes and metabolic pathways during AcD of WAS and MPW.

Synergistic effect and microbial community structure of waste-activated sludge and kitchen waste solids residue mesophilic anaerobic co-digestion.

Anaerobic co-digestion was conducted on the solid residues after three-phase separation of kitchen waste (KWS) and waste-activated sludge (WAS), the synergistic effects and process performance were studied during co-digestion at different ratios of KWS to WAS. KWS and WAS mix ratios of 0:1, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1 and 1:0 (based on TS). The results showed that a ratio of KWS to WAS of 1:1 got a very high methane recovery with a methane yield of 310.45 ± 30.05 mL/g VSadded. The highest concentration of free ammonia among all reaction systems was only 70.23 ± 5.53 mg/L, which was not enough to produce ammonia inhibition in the anaerobic co-digestion system. However, when the KWS content exceeded 50%, methane inhibition and prolongation of the lag phase were observed due to the accumulation of volatile fatty acids (VFAs), and during the lag phase. Microbial community analysis showed that various bacterial groups involved in acid production and hydrolysis were mainly dominated by phylum Firmicutes, Chloroflexi, Proteobacteria and Bacteroidetes. Hydrogenotrophic methanogen was found to dominate all archaeal communities in the digesters. Co-digestion of KWS with WAS significantly increased the relative abundance of Methanobacterium compared with anaerobic digestion of WAS alone.

Source separation and anaerobic co-digestion of blackwater and food waste for biogas production and nutrient recovery.

Anaerobic co-digestion of source-separated blackwater (BW) and food and kitchen waste (FW) offers decentralized circular economy solutions by enabling local production of biogas and nutrient-rich byproducts. In this study, a 2 m3 pilot-scale continuously stirred tank reactor (CSTR) operated under mesophilic conditions was utilized for co-digestion of BW and FW. The process obtained a CH4 yield of 0.7 ± 0.2 m3/kg influent-volatile solid (VS), reaching a maximum yield of 1.1 ± 0.1 m3/kg influent-VS, with an average organic loading rate of 0.6 ± 0.1 kg-VS/m3/d and HRT of 25 days. The CH4 production rate averaged 0.4 ± 0.1 m3/m3/d, peaking at 0.6 ± 0.1 m3/m3/d. Treatment of digestate through flocculation followed by sedimentation recovered over 90% of ammonium nitrogen and potassium, and 80-85% of total phosphorus in the liquid fraction. This nutrient-rich liquid was used to cultivate Chlorella vulgaris, achieving a biomass concentration of 1.2 ± 0.1 g/L and 85 ± 3% and 78 ± 5% ammonium nitrogen and phosphorus removal efficiency, respectively. These findings not only highlight the feasibility of anaerobic co-digestion of source-separated BW and FW in local biogas production but also demonstrate the potential of microalgae cultivation as a sustainable approach to converting digestate into nutrient-rich algae biomass.

Optimization of ultrasonication and alkalinization as pretreatment methods for leachate co-digested with food waste toward maximum synergistic effects.

Anaerobic co-digestion (AcoD) of food waste (FW) and landfill leachate has shown promising results in enhancing the methane yield. However, leachate includes toxic and refractory compounds that may impact the decomposition process. In this research, co-digested leachate was pretreated using ultrasonication and alkalinization to manipulate its characteristics toward improved synergism with FW. Experimental optimization was conducted through biochemical methane potential (BMP) assays to identify the optimum operating conditions of the pretreatment methods. The study evaluated the synergistic effects of co-digestion with raw and pretreated leachate on enhancing the performance in terms of feedstock solubilization and methane production. The BMP test demonstrated that alkalinization and ultrasonication improved the total methane generation by 35% and 27%, respectively, yielding around 397 and 375 mL CH4 per g of volatile solids. Moreover, ultrasonication and alkalinization enhanced the synergistic effects by 28% and 36%, respectively, compared to co-digestion with untreated leachate. Optimization by response surface methodology revealed that maximum performance could be achieved with leachate sonication at 212 W for 37.5 min or augmenting 788 g NaOH per kg of volatile solids. Kinetic and statistical models were derived to simulate and assess the impacts of the pretreatment parameters on the AcoD process. The results indicated that the ultrasonication energy had a higher influence on total solubility and methane production than alkaline dosage. Additionally, energy efficiency analyses were performed to examine the overall viability of the examined management approach and found that alkalinization increased the net energy efficiency by 23%, whereas ultrasonication was inefficient within the examined laboratory conditions despite the improved performance. The findings support an integrated organic waste management system where separated FW is co-treated with landfill leachate.

Valorization of crop residues and animal wastes: Anaerobic co-digestion technology.

To switch the over-reliance on fossil-based resources, curb environmental quality deterioration, and promote the use of renewable fuels, much attention has recently been directed toward the implementation of sustainable and environmentally benign 'waste-to-energy' technology exploiting a clean, inexhaustible, carbon-neutral, and renewable energy source, namely agricultural biomass. From this perspective, anaerobic co-digestion (AcoD) technology emerges as a potent and plausible approach to attain sustainable energy development, foster environmental sustainability, and, most importantly, circumvent the key challenges associated with mono-digestion. This review article provides a comprehensive overview of AcoD as a biochemical valorization pathway of crop residues and livestock manure for biogas production. Furthermore, this manuscript aims to assess the different biotic and abiotic parameters affecting co-digestion efficiency and present recent advancements in pretreatment technologies designed to enhance feedstock biodegradability and conversion rate. It can be concluded that the substantial quantities of crop residues and animal waste generated annually from agricultural practices represent valuable bioenergy resources that can contribute to meeting global targets for affordable renewable energy. Nevertheless, extensive and multidisciplinary research is needed to evolve the industrial-scale implementation of AcoD technology of livestock waste and crop residues, particularly when a pretreatment phase is included, and bridge the gap between small-scale studies and real-world applications.

Incorporating saline microalgae biomass in anaerobic digester treating sewage sludge: Impact on performance and microbial populations.

The aim of this study was to acclimate anaerobic prokaryotes to saline microalgae biomass. Semi-continuous experiments were conducted using two 1.5 L mesophilic reactors for 10 weeks, (hydraulic retention time of 21 days). The first reactor was solely fed with sewage sludge (control), while the second received a mixture of sewage sludge and microalgal biomass (80/20 %w/w) cultivated at 70 g·L-1 salinity. The in-reactor salinity reached after the acclimation phase was 14 g·L-1. Biomethane production was comparable between the control and acclimated reactors (205 ± 29 NmLMethane·gVolatileSolids-1). Salinity tolerance assessment of methanogenic archaea revealed that salinity causing 50% inhibition of methane production increased from 10 to 27 g·L-1 after acclimation. Microbial diversity analyses revealed notable changes in methanogenic archaea populations during co-digestion of saline microalgae biomass, particularly methylotrophic (+27%) and acetotrophic (-26%) methanogens. This study has highlighted the possibility of treating efficiently saline microalgae in co-digestion with sewage sludge in future industrial biogas plants.

Fractional calculus as a generalized kinetic model for biochemical methane potential tests.

This study presents a fractional calculus model as a generalized kinetic model for estimating the maximum methane yield and degradation kinetics in biomethane potential (BMP) assays, a key analytical method in anaerobic digestion research and application. The fractional model outperformed common first-order kinetic models by yielding superior data fitting and properly managing substrate heterogeneity. The fractional model showed robust performance in mono-digestion, co-digestion and pre-treatment BMP assays with or without presence of large tailing or sigmoidal patterns in the BMP curve. The main advantage of the fractional model over other models is its ability to capture the complexities of the methane production process without losing model accuracy. Assessment of the mathematical model revealed that for fractional orders greater than 0.8 the Mittag-Leffler sequence could be transformed into a more computationally efficient exponential function.

Study on the Environmental Impact and Benefits of Incorporating Humus Composites in Anaerobic Co-Digestion Treatment.

This study evaluated the environmental impact and overall benefits of incorporating humus composites in the anaerobic co-digestion of kitchen waste and residual sludge. The life cycle assessment method was used to quantitatively analyze the environmental impact of the entire anaerobic co-digestion treatment process of waste, including garbage collection, transportation, and final product utilization. Moreover, the comprehensive assessment of the environmental impact, energy-saving and emission-reduction abilities, and economic cost of using humus composites in the anaerobic co-digestion treatment process was conducted using a benefit analysis method. The results showed that the anaerobic co-digestion of kitchen waste and residual sludge significantly contributed to the mitigation of global warming potential (GWP), reaching -19.76 kgCO2-eq, but had the least impact on the mitigation of acidification potential (AP), reaching -0.10 kgSO2-eq. In addition, the addition of humus composites significantly increased the production of biogas. At a concentration of 5 g/L, the biogas yield of the anaerobic co-digestion process was 70.76 m3, which increased by 50.62% compared with the blank group. This amount of biogas replaces ~50.52 kg of standard coal, reducing CO2 emissions by 13.74 kg compared with burning the same amount of standard coal. Therefore, the anaerobic co-digestion treatment of kitchen waste and residual sludge brings considerable environmental benefits.

Rapid characterization of sulfur and phosphorus in organic waste by near infrared spectroscopy.

Near-infrared spectroscopy (NIRS) has recently emerged as a valuable tool for monitoring organic waste utilized in anaerobic digestion processes. Over the past decade, NIRS has significantly improved the characterization of organic waste by enabling the prediction of several crucial parameters such as biochemical methane potential, carbohydrate, lipid and nitrogen contents, Chemical Oxygen Demand, and kinetic parameters. This study investigates the application of NIRS for predicting the levels of Sulfur (S) and Phosphorus (P) within organic waste materials. The results for sulfur prediction exhibited a high level of accuracy, yielding an error of 1.21 g/Kg[TS] in an independently validated dataset, coupled with an R-squared value of 0.84. Conversely, the prediction of phosphorus proved to be slightly less successful, showing an error of 1.49 g/Kg[TS] with an R-squared value of 0.70. Furthermore, the disparities in performance seem to stem from the inherent correlation between the spectral data and the sulfur or phosphorus contents. Significantly, a variable selection technique known as CovSel was employed, shedding light on the differing approaches used for sulfur and phosphorus predictions. In the case of sulfur, the prediction was achieved through a direct correlation with wavelengths associated with sulfur-related functional groups (such as R - S(=O)2 - OH, -SH, and R-S-S-R) present in the NIR spectra. In contrast, phosphorus prediction relied on an indirect correlation with absorption bands related to organic matter (including CH, CH2, CH3, -CHO, R-OH, C = O, -CO2H, and CONH).

Comparative study of high-performance mesophilic and thermophilic anaerobic membrane bioreactors in the co-digestion of sewage sludge and food waste: Methanogenic performance and energy recovery potential.

To support smart cities in terms of waste management and bioenergy recovery, the co-digestion of sewage sludge (SeS) and food waste (FW) was conducted by the anaerobic membrane bioreactor (AnMBR) under mesophilic and thermophilic conditions in this study. The biogas production rate of the thermophilic AnMBR (ThAnMBR) at the SeS to FW ratio of 0:100, 75:25, 50:50 and 100:0 was 2.84 ± 0.21, 2.51 ± 0.26, 1.54 ± 0.26 and 1.31 ± 0.08 L-biogas/L/d, inconspicuous compared with that of the mesophilic AnMBR (MeAnMBR) at 3.00 ± 0.25, 2.46 ± 0.30, 1.63 ± 0.23 and 1.30 ± 0.17 L-biogas/L/d, respectively. The higher hydrolysis ratio and the poorer rejection efficiencies of the membrane under thermophilic conditions, resulting that the permeate COD, carbohydrate and protein of the ThAnMBR was higher than that of the MeAnMBR. The lost COD that might be converted into biogas was discharged with the permeate in the ThAnMBR, which was partly responsible for the inconspicuous methanogenic performance. Furthermore, the results of energy recovery potential assessment showed that the energy return on investment (EROI) of the MeAnMBR was 4.54, 3.81, 2.69 and 2.22 at the four SeS ratios, which was higher than that of the ThAnMBR at 3.29, 2.97, 2.02 and 1.80, respectively, indicating the advantage of the MeAnMBR over the ThAnMBR in energy recovery potential. The outcomes of this study will help to choose a more favorable temperature to co-digest SeS and FW to support the construction of smart cities.

Environmental impact of sewage sludge co-digestion with food waste and fat-oil-grease: Integrating plant-wide modeling with life cycle assessment approach.

Anaerobic co-digestion of fat-oil-grease (FOG) and food waste (FW) with sewage sludge (SS) in wastewater treatment plants is a method used to increase biogas production. In this study, digestion scenarios were compared using plant-wide modeling and life cycle assessment: Scenario-0 (mono-digestion of waste-activated sludge (WAS)), Scenario-1 (co-digestion of WAS with FOG), and Scenario-2 (co-digestion of WAS with FW). Scenario-0, with the highest energy use and landfilling of FOG/FW, has the worst environmental impact. Scenario-1 and Scenario-2 minimize the environmental load by energy recovery and avoiding landfilling of organic waste. Scenario-wise, the change in greenhouse gas (GHG) emissions from treatment was negligible. However, due to the impact of landfilling, GHG emissions in Scenario-0 were 21% and 30% higher than in Scenario-1 and 2, respectively. The environmental benefit of anaerobic co-digestion of FOG/FW with SS is not only in the contribution to energy production but also in the recycling of organic waste.

Food waste impact on dry anaerobic digestion of straw in a novel reactor: Biogas yield, stability, and hydrolysis-methanogenesis processes.

Gradient anaerobic digestion reactor (GADR) can improve substrate utilization efficiency by solving the problem of the "short circuit" of materials. However, the substrate's composition significantly affects the reactor's performance. This study investigated the impact of food waste (FW) levels on corn straw's dry anaerobic digestion (AD) in a novel GADR. The results show that biomethane production can be improved by coupling urban and agricultural solid waste recycling. The mechanism is to increase the hydrolysis and acid production efficiency, and the abundance of enzymes related to methanogenesis. The maximum methane yield (494.2 mL CH4/g VS) and the highest anaerobic biodegradability (85.7 %) were obtained when the FW was added at 60 %. The co-digestion of FW and straw can improve the hydrolysis and acid production efficiency and methane yield, which improves the buffering capacity and stability of the system compared with the single digestion of FW.

Enhancing biomethanation performance through co-digestion of diverse organic wastes: a comprehensive study on substrate optimization, inoculum selection, and microbial community analysis.

A blend of organic municipal solid waste, slaughterhouse waste, fecal sludge, and landfill leachate was selected in different mixing ratios to formulate the best substrate mixture for biomethanation. Individual substrates were characterized, and the mixing ratio was optimized with the help of a response surface methodology tool to a value of 1:1:1:1 (with a C/N ratio of 28±0.769 and total volatile fatty acid (VFA) concentration of 2500±10.53 mg/L) to improve the overall biomethanation. The optimized blend (C/N ratio: 28.6, VFA: 2538 mg/L) was characterized for physicochemical, biological, and microbial properties and subjected to anaerobic digestion in lab-scale reactors of 1000 mL capacity with and without the addition of inoculum. The biogas yield of individual substrates and blends was ascertained separately. The observed cumulative biogas yield over 21 days from the non-inoculated substrates varied between 142±1.95 mL (24.6±0.3 ml/gVS) and 1974.5±21.72 mL (270.4±3.1 ml/gVS). In comparison, the addition of external inoculation at a 5% rate (w/w) of the substrate uplifted the minimum and maximum cumulative gas yield values to 203±9.9 mL (35.0±1.6 mL/gVS) and 3394±13.4 mL (315.3±1.2 mL/gVS), respectively. The inoculum procured from the Defence Research and Development Organisation (DRDO) was screened in advance, considering factors such as maximizing VFA production and consumption rate, biogas yield, and digestate quality. A similar outcome regarding biogas yield and digestate quality was observed for the equivalent blend. The cumulative gas yield increased from 2673±14.5 mL (373.7±2.2 mL/gVS) to 4284±111.02 mL (391.47±20.02 mL/gVS) over 21 days post-application of a similar dosage of DRDO inoculum. The 16S rRNA genomic analysis revealed that the predominant bacterial population belonged to the phylum Firmicutes, with the majority falling within the orders Clostridiales and Lactobacillales. Ultimately, the study advocates the potential of the blend mentioned above for biomethanation and concomitant enrichment of both biogas yield and digestate quality.

Synergistic integration of hydrothermal pretreatment and co-digestion for enhanced biogas production from empty fruit bunches in high solids anaerobic digestion.

This study investigates the co-digestion of hydrothermally pretreated empty fruit bunches (EFB) at 190 °C for 5 min (HTP190-EFB) with decanter cake (DC) to improve biogas production in high solid anaerobic digestion (HSAD). The HTP190-EFB exhibited a 67.98 % reduction in total solids, along with the production of 0.89 g/L of sugar, 2.39 g/L of VFA, and 0.56 g/L of furfural in the liquid fraction. Co-digestion of HTP190-EFB with DC at mixing ratios of 5, 10, and 15 %w/v demonstrated improved methane yields and process stability compared to mono-digestion of HTP190-EFB. The highest methane yield of 372.69 mL CH4/g-VS was achieved in the co-digestion with 5 %w/v DC, representing a 15 % increase compared to digestion of HTP190-EFB (324.30 mL CH4/g-VS) alone. Synergistic effects were quantified, with the highest synergistic methane yield of 77.65 mL CH4/g-VS observed in the co-digestion with 5 %w/v DC. Microbial community analysis revealed that co-digestion of hydrothermally pretreated EFB with decanter cake promoted the growth of Clostridium sp., Lactobacillus sp., Fibrobacter sp., Methanoculleus sp., and Methanosarcina sp., contributing to enhanced biogas production compared to mono-digestion of pretreated EFB. Energy balance analysis revealed that co-digestion of HTP190-EFB with DC resulted in a total net energy of 599.95 kW, 52 % higher than mono-digestion of HTP190-EFB (394.62 kW). Economic analysis showed a shorter return on investment for the co-digestion system (0.86 years) compared to the mono-digestion of HTP190-EFB (1.02 years) and raw EFB (2.69 years). The co-digestion of HTP190-EFB with 5 %w/v DC offers a promising approach to optimize methane yield, process stability, and economic feasibility, supporting the palm oil industry for producing renewable energy and sustainable waste management.

Improving anaerobic digestion performance after severe acidification: Unveiling the impacts of Fe3O4-bentonite composites in co-digestion of waste activated sludge and food waste.

Acidification recovery in anaerobic digestion of food waste is challenging. This study explored its in-situ recovery using a co-substrate of food waste and waste activated sludge. Fe3O4 and bentonite were used as conductor and carrier, respectively, to enhance AD performance under severe acidification. The application of Fe3O4-bentonite resulted in a 152% increase in cumulative methane in the Fe3O4-bentonite 10 digester, demonstrating its effectiveness in restoring the acidified AD system. In acidified systems, bentonite enhanced the diversity and richness of microbial communities due to its buffering capacity. The excessive non-conductive polysaccharides excreted by bacteria in extracellular polymeric substances reduced the possibility of electron transfer by Fe3O4. However, in the synergistic application of Fe3O4 and bentonite, this resistance was alleviated, increasing the possibility of direct interspecies electron transfer, and accelerating the consumption of volatile fatty acids. This approach of integrating carrier and conductive materials is significant for in-situ restoration of acidified systems.

Anaerobic co-digestion of human excreta, food leftovers and kitchen residue: 1 ternary mixture design, synergistic effects and RSM approach.

Anaerobic digestion of multiple substrates can generate more biogas while remaining stable, if positive synergistic effects are achieved. The type of co-digested substrates and the mixing ratio used, are the most important variables as each substrate has unique set of characteristics. Optimizing the volume ratios by testing various substrate mixing ratios is a popular method for determining the best-performing ratio of substrate mixture. The ternary mixture design has reportedly been found to quicken the process of testing different mixing ratios with high accuracy without running several experiments. Therefore, a ternary mixture design and a response surface approach are used in this work to ascertain the relationship between substrate mix and responses (biogas yield, methane yield, and synergy). The findings of the experiment revealed that R9 comprising 78.8 % human excreta, 11.8 % food leftovers and 9.4 % kitchen residue, had the highest methane production of 764.79 mLCH4/gVS and a synergistic index of 3.26. Additionally, the 3D response surface plots from the response surface model showed important and shared interactions between Human Excreta, (HE), Food Leftovers (FLO), and Kitchen Residue (KR). HE and KR had a similar positive synergistic effect on biogas yield, methane yield, and synergy, which was not the case for FLO. The response surface plots showed that the predicted responses (methane yield, biogas yield and synergy) increased with increasing HE and KR fractions and decreased with increasing FLO fractions in the substrate mixtures.