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As antibiotic resistance continues to spread globally, there is growing interest in the potential to limit the spread of antibiotic resistance genes (ARGs) from wastewater sources. In particular, operational conditions during sludge digestion may serve to discourage selection of resistant bacteria, reduce horizontal transfer of ARGs, and aid in hydrolysis of DNA. This study applied metagenomic analysis to examine the removal efficiency of ARGs through thermophilic and mesophilic anaerobic digestion using bench-scale reactors. Although the relative abundance of various ARGs shifted from influent to effluent sludge, there was no measureable change in the abundance of total ARGs or their diversity in either the thermophilic or mesophilic treatment. Among the 35 major ARG subtypes detected in feed sludge, substantial reductions (removal efficiency >90 %) of 8 and 13 ARGs were achieved by thermophilic and mesophilic digestion, respectively. However, resistance genes of aadA, macB, and sul1 were enriched during the thermophilic anaerobic digestion, while resistance genes of erythromycin esterase type I, sul1, and tetM were enriched during the mesophilic anaerobic digestion. Efflux pump remained to be the major antibiotic resistance mechanism in sludge samples, but the portion of ARGs encoding resistance via target modification increased in the anaerobically digested sludge relative to the feed. Metagenomic analysis provided insight into the potential for anaerobic digestion to mitigate a broad array of ARGs.

Up-flow anaerobic sludge blanket (UASB) reactor belongs to high-rate systems, able to perform anaerobic reaction at reduced hydraulic retention time, if compared to traditional digesters. In this review, the most recent advances in UASB reactor applications are critically summarized and discussed, with outline on the most critical aspects for further possible future developments. Beside traditional anaerobic treatment of soluble and biodegradable substrates, research is actually focusing on the treatment of refractory and slowly degradable matrices, thanks to an improved understanding of microbial community composition and reactor hydrodynamics, together with utilization of powerful modeling tools. Innovative approaches include the use of UASB reactor for nitrogen removal, as well as for hydrogen and volatile fatty acid production. Co-digestion of complementary substrates available in the same territory is being extensively studied to increase biogas yield and provide smooth continuous operations in a circular economy perspective. Particular importance is being given to decentralized treatment, able to provide electricity and heat to local users with possible integration with other renewable energies. Proper pre-treatment application increases biogas yield, while a successive post-treatment is needed to meet required effluent standards, also from a toxicological perspective. An increased full-scale application of UASB technology is desirable to achieve circular economy and sustainability scopes, with efficient biogas exploitation, fulfilling renewable energy targets and green-house gases emission reduction, in particular in tropical countries, where limited reactor heating is required.

The anaerobic digestion performance correlates with the functional microbial community. Mesophilic and thermophilic digestions of vegetable waste were conducted, and dynamics of the microbial community were investigated. The mesophilic and thermophilic collapsed stages occurred at organic loading rates of 1.5 and 2.0 g VS/(L d) due to the accumulation of volatile fatty acids with final concentrations of 2276 and 6476 mg/L, respectively. A high concentration of volatile fatty acids caused the severe inhibition of methanogens, which finally led to the imbalance between acetogenesis and methanogenesis. The mesophilic digestion exhibited a higher microbial diversity and richness than the thermophilic digestion. Syntrophic acetate-oxidizing coupled with hydrogenotrophic methanogenesis was the dominant pathway in the thermophilic stable system, and acetoclastic methanogenesis in the mesophilic stable system. The dominant acidogens, syntrophus, and methanogens were unclassified_f__Anaerolineaceae (8.68%), Candidatus_Cloacamonas (19.70%), Methanosaeta (6.10%), and Methanosarcina (4.08%) in the mesophilic stable stage, and Anaerobaculum (12.59%), Syntrophaceticus (4.84%), Methanosarcina (30.58%), and Methanothermobacter (3.17%) in thermophilic stable stage. Spirochaetae and Thermotogae phyla were the characteristic microorganisms in the mesophilic and thermophilic collapsed stages, respectively. These findings provided valuable information for the deep understanding of the difference of the microbial community and methane-producing mechanism between mesophilic and thermophilic digestion of vegetable waste.

Municipal solid waste contains mainly organic wastes that can be a good source for anaerobic digestion. Solid-state anaerobic digestion is an affordable and suitable technique to mitigate the organic fraction of municipal solid waste (OFMSW). However, as the organic loading of OFMSW is high, co-digestion with other materials can improve the system's performance. This study aimed to investigate the performance of the co-digestion of OFMSW and sawdust and study the parameters affecting its performance. Based on the experiments, the optimum sawdust/OFMSW ratio was achieved 1:2 with the methane production of 0.3 L/g VS. In addition, the inoculum-to-substrate ration (I/S) was investigated at 1:4, 1:2, 1:1, 2:1 ratios. The best result was obtained at 2:1 ratio with a total methane yield of 0.28 L/g VS. The results also indicated that I/S ratios less than 1:1 led to fatty acid accumulation and acidic pH condition. The effect of total solids content on the co-digestion process was also examined in this study. According to the results, as the total solids increased, the biomethane yield decreased while the biogas content increased.

Antibiotic resistance genes (ARGs) have emerged as a significant global health threat, contributing to fatalities worldwide. Wastewater treatment plants (WWTPs) and livestock farms serve as primary reservoirs for these genes due to the limited efficacy of existing treatment methods and microbial adaptation to environmental stressors. Anaerobic digestion (AD) stands as a prevalent biological treatment for managing sewage sludge and manure in these settings. Given the agricultural utility of AD digestate as biofertilizers, understanding ARGs’ fate within AD processes is essential to devise effective mitigation strategies. However, understanding the impact of various factors on ARGs occurrence, dissemination, and fate remains limited. This review article explores various AD treatment parameters and correlates to various resistance mechanisms and hotspots of ARGs in the environment. It further evaluates the dissemination and occurrence of ARGs in AD feedstocks and provides a comprehensive understanding of the fate of ARGs in AD systems. This review explores the influence of key AD parameters such as feedstock properties, pretreatments, additives, and operational strategies on ARGs. Results show that properties such as high solid content and optimum co-digestion ratios can enhance ARG removal, while the presence of heavy metals, microplastics, and antibiotics could elevate ARG abundance. Also, operational enhancements, such as employing two-stage digestion, have shown promise in improving ARG removal. However, certain pretreatment methods, like thermal hydrolysis, may exhibit a rebounding effect on ARG levels. Overall, this review systematically addresses current challenges and offers future perspectives associated with the fate of ARGs in AD systems. Graphical Abstract

Anaerobic digestion of fruit and vegetable waste (FVW) offers an environmentally friendly alternative for waste disposal, converting it into methane for energy recovery. Typically, FVW digestion is conducted in a continuously stirred tank reactor (CSTR) due to its ease of use and stability with solid concentrations between 5 and 10%. However, CSTRs are limited to organic loading rates (OLRs) of about 3 kg COD/m 3 .day, resulting in large reactor volumes, low methane productivity, and costly wet digestate handling. This work introduces a novel method for methane production from FVW using a high-rate reactor system. The proposed approach involves grinding, centrifuging, and/or pressing the FVW to separate it into liquid and solid phases. The liquid phase is then digested in an up-flow anaerobic sludge blanket (UASB) reactor, while the solid phase undergoes digestion in a dry methanization reactor. A model incorporating all biological reactors was implemented in the Anaerobic Digestion Model 1 (ADM1) to provide a theoretical basis for the experimental development of this system. The current simulation scenarios offer initial references for operating the experimental system, which will, in turn, generate data for further model refinement. For instance, constrained liquid–gas mass transfer was considered for dry fermentation, with additional potential biochemical kinetic limitations to be incorporated following on experimental evidence. The success of this system could enable energy recovery in 72 Central Wholesale Markets across Brazil, offering a critical tool for planning, operating, and optimizing such systems.

During advanced biological wastewater treatment, a huge amount of sludge is produced as a by-product of the treatment process. Hence, reuse and recovery of resources and energy from the sludge is a big technological challenge. The processing of sludge produced by Wastewater Treatment Plants (WWTPs) is massive, which takes up a big part of the overall operational costs. In this regard, anaerobic digestion (AD) of sewage sludge continues to be an attractive option to produce biogas that could contribute to the wastewater management cost reduction and foster the sustainability of those WWTPs. At the same time, AD reduces sludge amounts and that again contributes to the reduction of the sludge disposal costs. However, sludge volume minimization remains, a challenge thus improvement of dewatering efficiency is an inevitable part of WWTP operation. As a result, AD parameters could have significant impact on sludge properties. One of the most important operational parameters influencing the AD process is temperature. Consequently, the thermophilic and the mesophilic modes of sludge AD are compared for their pros and cons by many researchers. However, most comparisons are more focused on biogas yield, process speed and stability. Regarding the biogas yield, thermophilic sludge AD is preferred over the mesophilic one because of its faster biochemical reaction rate. Equally important but not studied sufficiently until now was the influence of temperature on the digestate quality, which is expressed mainly by the sludge dewateringability, and the reject water quality (chemical oxygen demand, ammonia nitrogen, and pH). In the field of comparison of thermophilic and mesophilic digestion process, few and often inconclusive research, unfortunately, has been published so far. Hence, recommendations for optimized technologies have not yet been done. The review presented provides a comparison of existing sludge AD technologies and the gaps that need to be filled so as to optimize the connection between the two systems. In addition, many other relevant AD process parameters, including sludge rheology, which need to be addressed, are also reviewed and presented.

A novel coupling process using an aerobic bacterial reactor with nitrification and sulfur–oxidization functions followed by a microalgal reactor was proposed for simultaneous biogas desulfurization and anaerobic digestion effluent (ADE) treatment. ADE nitrified by bacteria has a potential to be directly used as a culture medium for microalgae because ammonium nitrogen, including inhibitory free ammonia (NH3), has been converted to harmless NO3−. To demonstrate this hypothesis, Chlorella sorokiniana NIES-2173, which has ordinary NH3 tolerance; that is, 1.6 mM of EC50 compared with other species, was cultivated using untreated/treated ADE. Compared with the use of a synthetic medium, when using ADE with 1–10-fold dilutions, the specific growth rate and growth yield maximally decreased by 44% and 88%, respectively. In contrast, the algal growth using undiluted ADE treated by nitrification–desulfurization was almost the same as with using synthetic medium. It was also revealed that 50% of PO43− and most metal concentrations of ADE decreased following nitrification–desulfurization treatment. Moreover, upon NaOH addition for pH adjustment, the salinity increased to 0.66%. The decrease in metals mitigates the bioconcentration of toxic heavy metals from wastewater in microalgal biomass. Meanwhile, salt stress in microalgae and limiting nutrient supplementation, particularly for continuous cultivation, should be of concern.

This study applied Illumina high-throughput sequencing to explore the microbial communities and functions in anaerobic digestion sludge (ADS) from two wastewater treatment plants based on a metagenomic view. Taxonomic analysis using SILVA SSU database indicated that Proteobacteria (9.52–13.50 %), Bacteroidetes (7.18 %–10.65 %) and Firmicutes (7.53 %–9.46 %) were the most abundant phyla in the ADS. Differences of microbial communities between the two types of ADS were identified. Genera of Methanosaeta and Methanosarcina were the major methanogens. Functional analysis by SEED subsystems showed that the basic metabolic functions of metagenomes in the four ADS samples had no significant difference among them, but they were different from other microbial communities from activated sludge, human faeces, ocean and soil. Abundances of genes in methanogenesis pathway were also quantified using a methanogenesis genes database extracted from KEGG. Results showed that acetotrophic was the major methanogenic pathway in the anaerobic sludge digestion.

In this study, the influence of temperature on biogas production efficiency and the microbial community structure was investigated in a two-phase anaerobic digestion reactor for co-digestion of cow manure and corn straw. The results illustrated that the contents of solluted chemical oxygen demand (SCOD) and volatile fatty acid (VFA) in the acidogenic phase and biogas production in the methanogenic phase maintained relatively higher levels at temperatures ranging from 35–25 °C. The methane content of biogas production could be maintained higher than 50% at temperatures above 25 °C. The microbial community structure analysis indicated that the dominant functional bacteria were Acinetobacter, Acetitomaculum, and Bacillus in the acidogenic phase and Cenarchaeum in the methanogenic phase at 35–25 °C. However, the performances of the acidogenic phase and the methanogenic phase could be significantly decreased at a lower temperature of 20 °C, and microbial activity was inhibited obviously. Accordingly, a low temperature was adverse for the performance of the acidogenic and methanogenic phases, while moderate temperatures above 25 °C were more conducive to high biogas production efficiency.