The organic loading rate (OLR) is a critical parameter affecting the stability of dry anaerobic digestion (AD) of kitchen waste (KW), and significantly impacting the variations in physicochemical parameters and microbial communities. However, the evolution of quorum sensing (QS) and their role on anaerobic biochemical metabolism during the increase in OLR in dry AD remain unknown. Therefore, this study systematically elucidated the matter through multi-omics analysis based on a pilot-scale dry AD of KW. The results demonstrated that fluctuations in the OLR significantly influenced the microbial QS in dry AD. When the OLR ≤4.0 g·VS/L·d, the system operated stably, and methane production increased. The enrichment of Proteobacteria was crucial for sustaining high levels of functional genes associated with various types of QS, including acyl-homoserine lactones (AI-1), autoinducer-2 (AI-2), autoinducer-3 (AI-3), and gamma-aminobutyric acid (GABA). This enabled cooperative communication among microbes under low OLR. Furthermore, most genes associated with these QS processes positively affected hydrolysis, acidogenesis, and methanogenesis. When the OLR increased to 6.0 g·VS/L·d, the fatty acids and hydrogen partial pressure increased significantly. The autoinducing peptides (AIP)-type became the predominant QS and was positively correlated with fatty acids abundance. Syntrophaceticus and Syntrophomonas may promote syntrophic oxidation of acetate at high OLR through AIP-type QS. These findings provided new insights into the QS processes of microbes during dry AD of KW and a theoretical foundation for optimizing biochemical metabolic processes in dry AD through QS.

Organic loading rate (OLR) is crucial for determining the stability of dry anaerobic digestion (AD). Digestate recirculation contributes to reactor stability and enhances methane production. Nevertheless, the understanding of how OLR and digestate recirculation affect the abundance and diversity of antibiotics and antibiotic resistance genes (ARGs), as well as the mechanisms involved in the dissemination of ARGs, remains limited. This study thoroughly investigated this critical issue through a long-term pilot-scale experiment. The metabolome analyses revealed the enrichment of various antibiotics, such as aminoglycoside, tetracycline, and macrolide, under low OLR conditions (OLR ≤ 4.0 g·VS/L·d) and the reactor instability. Antibiotics abundance decreased by approximately 19.66-31.69 % during high OLR operation (OLR ≥ 6.0 g·VS/L·d) with digestate recirculation. The metagenome analyses demonstrated that although low OLR promoted reactor stability, it facilitated the proliferation of antibiotic-resistant bacteria, such as Pseudomonas, and triggered functional profiles related to ATP generation, oxidative stress response, EPS secretion, and cell membrane permeability, thereby facilitating horizontal gene transfer (HGT) of ARGs. However, under stable operation at an OLR of 6.0 g·VS/L·d, there was a decrease in ARGs abundance but a notable increase in human pathogenic bacteria (HPB) and mobile genetic elements (MGEs). Subsequently, during reactor instability, the abundance of ARGs and HPB increased. Notably, during digestate recirculation at OLR levels of 6.0 and 7.0 g·VS/L·d, the process attenuated the risk of ARGs spread by reducing the diversity of ARGs hosts, minimizing interactions among ARGs hosts, ARGs, and MGEs, and weakening functional profiles associated with HGT of ARGs. Overall, digestate recirculation aids in reducing the abundance of antibiotics and ARGs under high OLR conditions. These findings provide advanced insights into how OLR and digestate recirculation affect the occurrence patterns of antibiotics and ARGs in dry AD.

Levulinic acid (LA) is a polymer with a vast industrial application range and can be co-produced as a minor by-product during the biological production of polyhydroxyalkanoates (PHA). However, the influence of key parameters as tools for favouring the production of LA over PHA is still unclear. In this study, we investigated how several critical operational conditions, i.e., carbon-nitrogen ratio (C/N), organic loading rate (OLR) and airflow, can be optimised to favour LA accumulation over PHA production by a mixed microbial culture (MMC), using synthetic grape pomace (GP) hydrolysate as the substrate. The results showed that it was possible to direct the MMC towards LA accumulation instead of PHA. The maximum LA yield was 2.7 ± 0.2 g LA/(L·d) using a C/N of 35, an airflow of 5 L/min and an OLR of 4 g sCOD/(L·d). The OLR and, to a lesser extent, the C/N ratio were the main factors significantly and positively correlated with the biological synthesis of LA.

The Anaerobic Baffled Reactor (ABR) is an effective solution for landfill leachate treatment using an anaerobic fermentation process, which helps to reduce operating costs and sludge volume. To better understand the biological, chemical, and physical processes involved, especially when combining the ABR with an aerobic component, the study aimed to investigate the performance of an Anaerobic-Aerobic Hybrid Baffled Reactor (AABR) that includes an Anaerobic Filter (AF) for treating landfill leachate. This research utilized two glass reactors. The first reactor, designated as AABR-AF, consisted of six independent rectangular glass chambers arranged side by side. The third and sixth chamber designed for aerobic treatment and AF, respectively. The second reactor was used as a control reactor and did not include any aerobic chamber. The highest Removal Efficiencies (REs) for turbidity, COD, BOD, TP, TKN, nitrate, TOC, and TSS in the AABR-AF and ABR-AF were found to be (65.4% and 56.3%), (98.3% and 94.1%), (98.1% and 93.2%), (86.4% and 65%), (89.2% and 76.7%), (81.2% and 64.4%), (88.2% and 79.4%), and (72.4% and 68.5%), respectively. These optimal REs were achieved at an HRT of 48 h and an OLR of 10 kg/m3.d. Also, the highest and the lowest REs in Heavy Metals (HMs) were 89.57% for manganese in AABR-AF and 6.59% for nickel in ABR-AF, in an OLR of 10 kg/m3.d, respectively. The effective removal of Organic Matters (OMs) from landfill leachate using the AABR-AF and ABR-AF was found to be strongly influenced by HRT and OLR. The AABR-AF configuration, featuring a single aerobic chamber in the reactor, exhibited a higher efficiency in removing OMs compared to the ABR-AF configuration.

Anaerobic digestion (AD) of microalgae is an intriguing approach for bioenergy production. The scaling-up of AD presents a significant challenge due to the systematic efficiency losses related to process instabilities. To gain a comprehensive understanding of AD behavior, this study assessed a modified version of the anaerobic digestion model No1 (ADM1) + Contois kinetics to represent microalgae AD impacted by overloading. To this end, two new inhibition functions were implemented: inhibition by acetate for acidogenesis/acetogenesis and total volatile fatty acids for hydrolysis. This proposed ADM1 modification (including Contois kinetics) simulated AD behavior during the stable, disturbed and recovery periods, showing that the inhibition functions described in the original ADM1 cannot explain the AD performance under one of the most common perturbations at industrial scale (overloading). The findings underscore the importance of refining the inhibitions present in original ADM1 to better capture and predict the complexities of microalgae AD against overloading.

The feasibility of anaerobic co-digestion in semicontinuous mode of two major urban biowaste, food waste (FW) and garden and park waste (GPW) (75 % FW and 25 % GPW) as well as the co-digestion of FW with the process water originated from the hydrothermal carbonization of GPW (95 % FW and 5 % process water), both on a COD basis, has been assessed. The effect of varying organic loading rate (OLR) from 1.5 to 3.5 g COD/L·d on methane yield, gross energy recovery, and microbiome population was evaluated. For comparison, anaerobic digestion of FW was also conducted to determine the best strategy for sustainable biowaste management. This study showed an optimal OLR of 2.5 g COD/L·d. Acetic and propionic acid content increased as OLR raised for each condition studied, while methane yield decreased at the highest OLR tested indicating overloading of the system. The anaerobic co-digestion of FW and process water showed a 10 % increase on methane production compared to anaerobic digestion of FW (324 vs. 294 mL CH4 STP/L·d). Moreover, it enhances the process due to a greater abundance and diversity of hydrolytic and acidogenic bacteria belonging to Bacterioidota, Firmicutes, and Chloroflexi phyla, as well as promotes the hydrogenotrophic pathway under higher propionic concentrations which is not usually favoured for methane production. The integration of hydrothermal carbonization of GPW with the anaerobic co-digestion of 95 % FW and 5 % of process water results in the highest potential energy recovery and could be a good strategy for sustainable management of urban biowaste.

The two-stage anaerobic digestion (AD) for biohythane production is a sustainable solution, but it is sensitive to organic shock load that disrupts reactors and inhibits biohythane production. This study investigated biohythane production, reactor performance, and the possibility of post-failure restoration in a two-stage AD system designed for treating high-strength organic wastewater. Sudden shock load was applied by increasing the OLR threefold higher after reaching steady state phase. During shock load phase, hydrogen content, hydrogen yield and methane production rate (MPR) reached its peak values of 62.61 %, 1.641 mol H2/mol glucose, and 1.003 L CH4/L⋅d respectively before declining significantly. Interestingly, during the restorative phase, hydrogen production sharply declined to nearly zero, while methane production exhibited a resilience and reached its peak methane content of 52.2 %. The study successfully demonstrated the system\'s resilience to sudden shock load, ensuring stable methane production, while hydrogen production did not exhibit the same capability.

Anaerobic digestion of high solid-content wastewater is hindered by high organic loading rates (OLRs). Granular activated carbon (GAC) was reported to promote direct interspecies electron transfer (DIET) and enhance reactor performance. In this study, three up-flow anaerobic sludge blanket (UASB) reactors were supplied with GAC in different locations: bottom (R1), top (R2), and bottom+top (R3). The performances of three reactors at different OLRs treating high solid-content wastewater were evaluated. At a low OLR, the highest methane yield (74 ± 4 %, g CH4-COD/g TCOD) was detected when GAC was supplied at top of the UASB (R2). When a high OLR was applied, the UASB supplemented with GAC at both bottom and top (R3) achieved the highest methane yield (66 ± 2 %, g CH4-COD/g TCOD), whereas the UASB supplemented with GAC at the top (R2) failed. Further studies on spatial distributions of sludge stability, specific methanogenic activities (SMAs), and microbial communities demonstrated the different impacts of GAC location on reactor performance and sludge characteristics under different OLRs. This study highlights the significance of considering organic loading capacity treating high solid-content wastewater when choosing GAC-based UASB systems.

This study deals with the effect of different organic loading rates (OLRs) on the organic load removal and the productivity of methane, in a microbial electrolysis cell-anaerobic digestion (MEC-AD) system treating raw waste activated sludge (WAS). For comparison, two identical reactors, a control (AD) and a MEC-AD reactor were constructed. Both reactors operated for 131 days, during which different Organic Loading Rates (OLRs) were examined; 1.1, 1.7 and 2.9 gCOD/(L*d). The results showed that the MEC-AD reactor performed better, relative to the AD reactor, at high OLRs. Specifically, the additional total particulate carbon and Kjeldahl nitrogen removal reached 12% and 13%, respectively, at an OLR of 1.7 gCOD/(L*d), while they reached 19% and 13%, respectively, at an OLR of 2.9 gCOD/(L*d). Moreover, the biogas production and the methane content increased by 30% and 6%, respectively, at an OLR of 1.7 gCOD/(L*d) and by 32% and 5%, respectively, at an OLR of 2.9 gCOD/(L*d). The electrochemical measurements indicated that the power output increased from 5 to 30 mW/m2 when the OLR increased from 1.1 to 2.9 gCOD/(L*d). Overall, the results showed that the MEC-AD accelerated and enhanced the WAS treatment, boosting methane productivity.

The semi-continuous anaerobic co-digestion (AcoD) of thermal and thermal-alkali pretreated organic fraction of municipal solid waste (OFMSW) and sewage sludge (SS) was studied under varying hydraulic retention times (HRT) and organic loading rates (OLR Three semi-continuous digesters were operated under control (non-pre-treated), thermally pretreated (125 °C), and thermal-alkali pretreated (125°C-3g/L NaOH) conditions at variable OLRs at 2.5, 4.0, 5.1, and 7.6 kgVS/m3.d and corresponding HRTs of 30, 20, 15, and 10 days. The 10 and 43% higher methane yield (0.445 m3/kgVS) and 11 and 57% higher VS removal (52%) was achieved for thermal-alkali pretreated digester at 5.1 kgVS/m3.d OLR over thermally pretreated (0.408 m3/kgVS, 45% VS removal) and control digesters (0.310 m3/kgVS, 33% VS removal), respectively. Thermal and thermal-alkali digesters failed on increasing the OLR to 7.6 kgVS/m3.d, whereas the control digester becomes upset at 5.1 kgVS/m3.d OLR. The metagenomic study revealed that Firmicutes, Bacteroidetes, Chloroflexi, Euryarchaeota, Proteobacteria, and Actinobacteria were the predominant bacterial population, whereas Methanosarcina and Methanothrix dominated the archaeal community. Energy balance analysis revealed that thermal alkali pretreatment showed the highest positive energy balance of 114.6 MJ/ton with an energy ratio of 1.25 compared with thermally pretreated (81.5 MJ/ton) and control samples (-46.9 MJ/ton). This work pave the way for scaleup of both thermal and thermal-alkali pre-treatment at 125 °C to realize the techno-economic and energy potential of the process.