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Most waste sludge belongs to long sludge-retention-time (SRT) sludge in China, but its outlet is becoming a severe environmental problem nowadays. In this study, the characteristics of mesophilic and thermophilic anaerobic digestion of long SRT sludge was investigated under different total solid content (TSC) of 4%, 6%, and 8% in batch reactors. The results showed that the methane production potential (Po) and methane production rate (Rmax) of thermophilic anaerobic digestion (TAD) were increased by about 60% and 200% compared with that of the mesophilic anaerobic digestion (MAD) at 4% TSC. In TAD groups, the Po of 240.95 LCH4 kgVS-1added and Rmax of 22.17 LCH4 kgVS-1added day-1 were achieved in the reactor of 4% TSC while the inhibition concentration of propionate and free ammonia reached 1022 mg L-1 and 603 mg L-1 in the reactor of 8% TSC. Although the diversity of microbial communities were poor in TAD, Coprothermobacter was enriched in TAD, which could degrade protein efficiently. In addition, the dewaterability of digested sludge in TAD was poorer than that in MAD, and both got worse with the increased TSC. Therefore, it is appropriate for the long SRT sludge to anaerobic digestion under thermophilic condition and lower TSC. So, thermophilic anaerobic digestion with lower TSC might be an alternative approach for the treatment and resource utilization of long SRT sludge.

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. [ABSTRACT FROM AUTHOR]

Polyacrylamide (PAM) is generally employed in wastewater treatment processes such as sludge dewatering and therefore exists in the sludge. Furthermore, it degrades slowly and can deteriorate methane yield during anaerobic digestion (AD). The impact or fate of PAM in AD under thermophilic conditions is still unclear. This study mainly focuses on PAM degradation and enhanced methane production from PAM-added sludge during 15 days of thermophilic (55°C) AD compared to mesophilic (35°C) AD. Sludge and PAM dose from 10 to 50 g/kg TSS were used. The results showed that PAM degraded by 76% to 78% with acrylamide (AM) content of 0.2 to 3.3 mg/L in thermophilic AD. However, it degraded only 27% to 30% with AM content of 0.5 to 7.2 mg/L in mesophilic AD. The methane yield was almost 230 to 238.4 mL g VSS on the 8th day in thermophilic AD but was 115.2 to 128.6 mL/g VSS in mesophilic AD. Mechanism investigation revealed that thermophilic AD with continuous stirring not only enhanced PAM degradation but also boosted the organics release from the sludge with added PAM and gave higher methane yield than mesophilic AD. [ABSTRACT FROM AUTHOR]

• Swine manure anaerobic digestion (AD) at 55 °C raised 47.9% methane than at 35 °C. • More oxytetracycline (OTC) led to less methane from mesophilic and thermophilic ADs. • Adding 400 mg/L OTC reduced energy conversion efficiencies of ADs by 21.4–48.4%. • More than 63.0% and 72.5% OTC were removed via mesophilic and thermophilic ADs. • Methanolinea was bearable to high content OTC in mesophilic and thermophilic ADs. Anaerobic digestion (AD) technology is a valuable method for producing biogas fuels and treating livestock wastes such as swine manures concurrently. However, the effect of emerging antibiotics on the AD process is still undiscovered. In this study, the influence of oxytetracycline (OTC) on the AD process was investigated under mesophilic (35 ± 0.5 °C) and thermophilic (55 ± 0.5 °C) conditions, respectively. The presence of OTC significantly inhibited the production of methane in AD process, where the methane yields decreased by 58.6% and 73.3% in mesophilic and thermophilic ADs when the initial concentration of OTC was 400 mg/L, respectively. Besides, OTC can be markedly degraded by the AD process with a removal efficiency higher than 90% when the OTC initial concentration is lower than 10 mg/L. Furthermore, a higher concentration OTC led to a lower biomethane yield, energy conversion efficiency, and contaminant removal during both mesophilic and thermophilic ADs. With adding of 400 mg/L OTC, Clostridium sensu stricto 1 (32.9%) and Anaerolinea (29.3%) are dominant to biodegrade organic matter during mesophilic and thermophilic AD systems. Correspondingly, Methanosaeta was functional in producing biomethane in both mesophilic (60.8%) and thermophilic (56.4%) AD systems. Additionally, Methanolinea was bearable to high concentrations of OTC during mesophilic and thermophilic AD processes. [ABSTRACT FROM AUTHOR]

This study presents a multi-criteria evaluation of energy recovery from wastewater sludges by the traditional anaerobic digestion pathway versus the emergent hydrothermal liquefaction (HTL) pathway. The two pathways were assessed considering the following 12 criteria: process stability, volatile solids reduction, process time, energy recovery, energy yield, net energy yield, energy return on investment, net power yield, energy density of the resulting biofuel, potential for methane emissions, effluent quality, and technology readiness. Experimental results from HTL of primary and secondary sludges and literature data on mesophilic and thermophilic anaerobic digestion are used here to quantify the above criteria for assessment. Although the two anaerobic pathways ranked better than the HTL pathway on the basis of energy return on investment (5.5–19.8 vs. 1.5), when all the 12 criteria were taken into consideration by the multi-criteria decision making approach, the HTL pathway outranked both the anaerobic pathways with a preference ranking of + 0.44 compared to − 0.28 in anaerobic digestion. It is recommended that developing technologies for recovering energy from the solid residue of HTL, the biochar, the energy return on investment by the HTL pathway could be improved from 1.5 to 2.42. • Evaluated energy recovery from sewage sludges by hydrothermal liquefaction (HTL). • Derived 12 performance criteria to compare HTL vs. anaerobic digestion (AD). • Used multicriteria decision-making (MCDM) method to compare HTL vs. AD. • MCDM method ranked HTL higher than both mesophilic and thermophilic AD. [ABSTRACT FROM AUTHOR]

Understanding fate of antibiotic resistant bacteria (ARB) vs. their antibiotic resistance genes (ARGs) during wastewater sludge treatment is critical in order to reduce the spread of antibiotic resistance through process optimization. Here, we spiked high concentrations of tetracycline-resistant bacteria, isolated from mesophilic (Iso M1-1--a Pseudomonas sp.) and thermophilic (Iso T10--a Bacillus sp.) anaerobic digested sludge, into batch digesters and monitored their fate by plate counts and quantitative polymerase chain reaction (QPCR) of their corresponding tetracycline ARGs. In batch studies, spiked ARB plate counts returned to baseline (thermophilic) or 1-log above baseline (mesophilic) while levels of the ARG present in the spiked isolate [tet(G)] remained high in mesophilic batch reactors. To compare results under semi-continuous flow conditions with natural influent variation, tet(O), tet(W), and sul1 ARGs, along with the intI1 integrase gene, were monitored over a 9-month period in the raw feed sludge and effluent sludge of lab-scale thermophilic and mesophilic anaerobic digesters. sul1 and intI1 in mesophilic and thermophilic digesters correlated positively (Spearman rho = 0.457-0.829, P < 0.05) with the raw feed sludge. There was no correlation in tet(O) or tet(W) ratios in raw sludge and mesophilic digested sludge or thermophilic digested sludge (Spearman rho = 0.130-0.486, P = 0.075-0.612). However, in the thermophilic digester, the tet(O) and tet(W) ratios remained consistently low over the entire monitoring period. We conclude that the influent sludge microbial composition can influence the ARG content of a digester, apparently as a result of differential survival or death of ARBs or horizontal gene transfer of genes between raw sludge ARBs and the digester microbial community. Notably, mesophilic digestion was more susceptible to ARG intrusion than thermophilic digestion, which may be attributed to a higher rate of ARB survival and/or horizontal gene transfer between raw sludge bacteria and the digester microbial community. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Thermophilic anaerobic digestion is a suitable technology to treat agricultural waste because of its higher biogas production, hygienisation effect and solids removal efficiency. Although these benefits, poor effluent quality and instability are encountered in some cases. The anaerobic co-digestion of winery waste and waste activated sludge at 55°C, operating at 23 days HRT and with an organic loading rate of 3.2 kg COD/m³d, was characterized by accumulation of volatile fatty acids, pH fall and reduction of biogas production, while mesophilic process was steady at long term. The study evaluated the effect of trace elements (iron, cobalt and nickel) augmentation in the thermophilic reactor at different concentration of micro-nutrients. The addition improved the process stability: pH became constant and average volatile fatty acids concentration was below 1,000 mg COD/L. The biogas production increased from 0.38 to 0.45 m³/kg COD, corresponding to 90% of COD removal, while mesophilic reactor removed the 78% of total COD. Digestate had interesting characteristics as fertilizer in fact the higher solids removal (28%) allowed to concentrate the phosphorus in the particulate fraction and nitrogen was transformed into more available form for plants growth. [ABSTRACT FROM AUTHOR]

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.

[Display omitted] • iARGs accounted for most ARGs during anaerobic digestion of cow manure. • AD increased absolute abundance of eARGs and bARGs. • Potential hosts of iARGs and eARGs were similar. • bARGs may spread between bacteria or across bacterial genera. • eARGs and bARGs were directly driven by physicochemical factors (e.g., pH). The fate of intracellular antibiotic resistance genes (iARGs), extracellular ARGs (eARGs) and bacteriophage ARGs (bARGs) during anaerobic digestion (AD) of cow manure is unclear. Thus, the characteristics of iARGs, eARGs and bARGs during mesophilic AD (MAD) and thermophilic AD (TAD) of cow manure were investigated. The absolute abundances of iARGs decreased by 69.82% after TAD. After MAD and TAD, the total absolute abundances of eARGs increased by 63.5 times and 67.6 times, respectively, whereas those of the bARGs increased by 47.60% and 59.22%. eARGs were mainly derived from the non-specific lysis of Firmicutes, Bacteroidetes, while bacteriophages had a wide range of hosts. The variations in iARGs, eARGs and bARGs were affected by the microbial hosts but also directly driven by physicochemical factors (e.g., pH). Overall, the findings of this study revealed that there may be a risk of eARGs and bARGs disseminating during the AD of cow manure. [ABSTRACT FROM AUTHOR]

Water resource recovery facilities have been described as creating breeding ground conditions for the selection, transfer, and dissemination of antibiotic resistance genes (ARGs) among various bacteria. The objective of this study was to determine the effect of direct addition of antibiotic and silver nanoparticles (Ag NPs, or nanosilver) on the occurrence of ARGs in thermophilic anaerobic digesters. Test thermophilic digesters were amended with environmentally-relevant concentrations of Ag NP (0.01, 0.1, and 1.0 mg-Ag/L; corresponding to ≈0.7, 7.0, and 70 mg-Ag/kg total solids) and sulfamethoxazole (SMX) that span susceptible to resistant classifications (1, 5, and 50 mg/L) as potential selection pressures for ARGs. Tetracycline (tet(O), tet(W)) and sulfonamide (sulI, sulII) ARGs and the integrase enzyme gene (/«ill) associated with Class 1 integrons were measured in raw sludge, test thermophilic digesters, a control thermophilic digester, and a control mesophilic digester. There was no apparent effect of Ag NPs on thermophilic anaerobic digester performance. The maximum SMX addition (50 mg/L) resulted in accumulation of volatile fatty acids and low pH, alkalinity, and volatile solids reduction. There was no significant difference between ARG gene copy numbers (absolute or normalized to 16S rRNA genes) in amended thermophilic digesters and the control thermophilic digester. Antibiotic resistance gene copy numbers in digested sludge ranged from 10³ to 106 copies per pL (≈8 X 10¹ to 8 X 104 copies per pg) of sludge as result of a 1-log reduction of ARGs (2-log reduction for mill). Quantities of the five ARGs in raw sludge ranged from 104 to 108 copies per pL (≈4 X 10² to 4 X 106 per pg) of sludge. Test and control thermophilic digesters (53 °C, 12-day solids retention time [SRT]) consistently reduced but did not eliminate levels of all analyzed genes. The mesophilic digester (37 °C, 20-day SRT) also reduced levels of sulI, sulll, and /«til genes, but levels of tet{O) and tet(W) were the same or higher than in raw sludge. Antibiotic resistance gene reductions remained constant despite the application of selection pressures, which suggests that digester operating conditions are a strong governing factor of the bacterial community composition and thus the prevalence of ARGs. [ABSTRACT FROM AUTHOR]

In this work, the impact of granular activated carbon (GAC) on the mesophilic and thermophilic anaerobic digestion of chicken manure and the structure of microbial communities was investigated. These results demonstrated that GAC supplementation effectively enhanced the consumption of produced organic acids in the mesophilic and thermophilic batch tests, accompanied by faster biomethane production in the presence of GAC than from reactors without GAC. However, since the free ammonia level was 3–6 times higher in the thermophilic reactors, this led to the instability of the anaerobic digestion process of the nitrogen-rich substrate at thermophilic temperatures. Bacteroidia and Clostridia were the two main bacterial classes in the mesophilic reactors, whereas the class Clostridia had a competitive advantage over other groups in the thermophilic systems. The archaeal communities in the mesophilic reactors were mainly represented by representatives of the genera Methanosarcina, Methanobacterium, and Methanotrix, whereas the archaeal communities in the thermophilic reactors were mainly represented by members of the genera Methanosarcina, Methanoculleus, and Methanothermobacter. New data obtained in this research will help control and manage biogas reactors in the presence of GAC at different temperatures. [ABSTRACT FROM AUTHOR]

The potential of a mesophilic-thermophilic-mesophilic anaerobic digestion system was investigated with respect to improvement of both digestion and sanitation efficiencies during treatment of liquid cattle manure. The pilot plant produced a high methane yield from liquid dairy cattle manure of 0.24 m³ (kg VSfed)-1. Considering the low system loading rate of 1.4–1.5 kg VS (m³ d)-1, digestion efficiency compared to conventional processes did not appear improved. The minimum guaranteed retention time in the tubular thermophilic reactor was increased compared to a continuously stirred tank reactor. Levels of intestinal enterococci in raw liquid manure as determined with cultivation methods were reduced by 2.5–3 log units to a level of around 10² cfu/mL. This sanitizing effect was achieved both during mesophilic-thermophilic-mesophilic and thermophilic-mesophilic treatment, provided the thermophilic digester was operated at 53–55°C. A change in feeding interval from 1 h to 4 h did not significantly alter methane yield and sanitation efficiency. It was proposed that a two-stage, thermophilic-mesophilic anaerobic digestion system would be able to achieve the same sanitizing effect and equal or better digestion efficiency at lower costs. [ABSTRACT FROM AUTHOR]

The fate of diverse antibiotic resistance genes (ARGs) in anaerobic digestion (AD) system under different temperature remains largely unknow. This study employed the shotgun metagenomic sequencing and 16S rRNA amplicon sequencing approaches and investigated the shifts of composition and abundance of more than 1360 ARGs, as well as microbial community, in a temperature-raising AD system (from 35 °C to 55 °C) treating municipal sewage sludge. Results suggested that the six ARGs, inlcuding mac B, tet A(58), bcr A, evg S, mtr A, and ole C, were predominated in digested sludge. More than 407ARG subtypes (30%) could be largely reduced under high temperature condition. Thermophilic AD (55 °C) provided a better removal for tetracycline, macrolide, penam, fluoroquinolone, acridine dye, and peptide resistance genes when compared to mesophilic AD (35 °C). The removal rates of these genes reached 29.7%–32.3%. In addition, at the end of thermophilic digestion process, the relative abundance of Firmicutes increased (from 18.22% to 74.89%), while Proteobacteria and Chloroflexi both decreased, from 36.39% to 6.80% and 20.29%–2.64%. This study further proposed the underlying mechanisms of effective elimination of ARGs under high temperature by: (1) the reduction of genes encoding antibiotic efflux pump, (2) the promotion of sludge biomass degradation rate, (3) the loss of microbial community diversity in AD systems, and (4) the inactivation of multidrug-resistant bacteria. The significance of current study is in characterizing the fate of ARGs across AD process, which allows to correlate diverse genetic properties with digesters' operational condition. Results provided an insight into the manipulation of AD technology (e.g. temperature) to efficiently remove ARGs from municipal sewage sludge. • Fate of ARGs in a temperature-raising anaerobic digester was investigated. • More than 1360 ARG subtypes were identified by metagenomics approach. • Effective reduction of ARGs under thermophilic condition was confirmed. • Underlying mechanisms of ARGs removal were proposed. [ABSTRACT FROM AUTHOR]

• Algal biochar enhanced co-digestion of algal biomass and food waste for biomethane. • 75% algal biomass + 25% food waste mixture co-digestion showed a synergistic effect. • Average methane yields enhanced by 12–54% due to 15 g/L of algal biochar amendment. • Proteiniphilum enriched in mesophilic reactor like Defluviitoga in thermophilic one. • Biochar shifted methanogens towards a balance of two flexible methanogenic pathways. The impact of algal biochar addition on mesophilic and thermophilic anaerobic co-digestion of algal biomass and food waste was investigated with a focus on semi-continuous operations and functional microbial communities. Under batch co-digestion, the highest co-digestion synergy was observed for a mixture of 25% food waste and 75% algal biomass. During semi-continuous co-digestion of 25% food waste-75% algal biomass mixture, biochar amended digesters exhibited a 12–54% increase in average methane yield (275.8–394.6 mL/gVS) compared to the controls. Elevated temperature induced narrow distributions of volatile fatty acids (VFAs) by inhibiting the production of branched VFAs. Genus Proteiniphilum was selectively enriched by 3.2 folds in mesophilic digesters with biochar amendment while genus Defluviitoga was selectively enriched in thermophilic digesters due to elevated temperature. Methanogenic communities were significantly different in mesophilic and thermophilic digesters. Biochar amendment contributed to shifts in the predominant methanogens leading to a more balanced state of two methanogenic pathways. [ABSTRACT FROM AUTHOR]