Anaerobic Fermentation Of Waste Activated Sludge Research Articles

Tannic Acid Modulation of Substrate Utilization, Microbial Community, and Metabolic Traits in Sludge Anaerobic Fermentation for Volatile Fatty Acid Promotion.

Anaerobic fermentation is a crucial route to realize effective waste activated sludge (WAS) resource recovery and utilization, while the overall efficiency is commonly restrained by undesirable disruptors (i.e., chemical dewatering agents). This work unveiled the unexpectedly positive effects of biodewatering tannic acid (TA) on the volatile fatty acids (VFAs) biosynthesis during WAS anaerobic fermentation. The total VFAs yield was remarkably increased by 15.6 folds with enriched acetate and butyrate in TA-occurred systems. TA was capable to disintegrate extracellular polymeric substances to promote the overall organics release. However, TA further modulated the soluble proteins structure by hydrogen bonding and hydrophobic interactions, resulting in the decrease of proteins bioavailability and consequential alteration of metabolic substrate feature. These changes reshaped the microbial community and stimulated adaptive regulatory systems in hydrolytic-acidogenic bacteria. The keystone species for carbohydrate metabolism (i.e., Solobacterium and Erysipelotrichaceae) were preferentially enriched. Also, the typical quorum sensing (i.e., enhancing substrate transport) and two-component systems (i.e., sustaining high metabolic activity) were activated to promote the microbial networks connectivity and ecological cooperative behaviors in response to TA stress. Additionally, the metabolic functions responsible for carbohydrate hydrolysis, transmembrane transport, and intracellular metabolism as well as VFA biosynthesis showed increased relative abundance, which maintained high microbial activities for VFAs biosynthesis. This study underscored the advantages of biodewatering TA for WAS treatment in the context of resource recovery and deciphered the interactive mechanisms.

Facilitating effects of the synergy with zero-valent iron and peroxysulfate on the sludge anaerobic fermentation system: Combined biological enzyme, microbial community and fermentation mechanism assessment

This study evaluated a synergetic waste activated sludge treatment strategy with environmentally friendly zero-valent iron nanoparticles (Fe0) and peroxysulfate. To verify the feasibility of the synergistic treatment, Fe0, peroxysulfate, and the mixture of peroxysulfate and Fe0 (synergy treatment) were added to different sludge fermentation systems. The study demonstrated that the synergy treatment fermentation system displayed remarkable hydrolysis performance with 435.50 mg COD/L of protein and 197.67 mg COD/L of polysaccharide, which increased 1.13–2.85 times (protein) and 1.12–1.49 times (polysaccharide) for other three fermentation system. Additionally, the synergy treatment fermentation system (754.52 mg COD/L) exhibited a well acidification performance which was 1.35–41.73 times for other systems (18.08–557.27 mg COD/L). The synergy treatment fermentation system had a facilitating effect on the activity of protease, dehydrogenase, and alkaline phosphatase, which guaranteed the transformation of organic matter. Results also indicated that Comamonas, Soehngenia, Pseudomonas, and Fusibacter were enriched in synergy treatment, which was beneficial to produce SCFAs. The activation of Fe0 on peroxysulfate promoting electron transfer, improving the active groups, and increasing the enrichment of functional microorganisms showed the advanced nature of synergy treatment. These results proved the feasibility of synergy treatment with Fe0 and peroxysulfate to enhance waste activated sludge anaerobic fermentation.

Thiosulfate/FeCl3 pre-treatment enhances short-chain fatty acid production and mitigates H2S generation during anaerobic fermentation of waste activated sludge: Performance, microbial community and ecological analyses

Anaerobic fermentation (AF) has been identified as a promising method of transforming waste activated sludge (WAS) into high-value products (e.g., short-chain fatty acids (SCFAs)). This study developed thiosulfate/FeCl3 pre-treatment and investigated the effects of different thiosulfate/FeCl3 ratios (S:Fe = 3:1, 3:2, 1:1, 3:4 and 3:5) on SCFA production and sulfur transformation during the AF of WAS. At a S:Fe ratio of 1:1, the maximal SCFA yield (933.3 mg COD/L) and efficient H2S removal (96.5 %) were obtained. S:Fe ratios ≤ 1:1 not only benefited hydrolysis and acidification but largely mitigated H2S generation. These results were supported by the enriched acidogens and reduced sulfur-reducing bacteria (SRB). Molecular ecological network analysis further revealed that the keystone taxon (g_Saccharimonadales) was found in S:Fe = 1:1, together with reductions in associations among methanogens, acidogens and SRB. This work provides a strategy for enhancing high-value product recovery from WAS and minimising H2S emissions.

Dynamic microbiome disassembly and evolution induced by antimicrobial methylisothiazolinone in sludge anaerobic fermentation for volatile fatty acids generation

In the post-COVID-19 pandemic era, various antimicrobials have emerged and concentrated in waste-activated sludge (WAS), affecting the biological treatment of WAS. However, there is still a knowledge gap in the dynamic response and adaptive mechanism of anaerobic microbiome under exogenous antimicrobial stress. This study found that methylisothiazolinone (MIT, as a typic antimicrobial) caused an interesting lag effect on the volatile fatty acids (VFAs) promotion in the WAS anaerobic fermentation process. MIT was effective to disintegrate the extracellular polymeric substances (EPS), and those functional anaerobic microorganisms were easily exposed and negatively impacted by the MIT interference after the loss of protective barriers. Correspondingly, the ecological interactions and microbial metabolic functions related to VFA biosynthesis (e.g., pyruvate metabolism) were downregulated at the initial stage. The syntrophic consortia gradually adapted to the interference and attenuated the MIT stress by activating chemotaxis and resistance genes (e.g., excreting, binding, and inactivating). Due to the increased bioavailable substrates in the fermentation systems, the dominant microorganisms (i.e., Clostridium and Caloramator) with both VFAs production and MIT-tolerance functions have been domesticated. Moreover, MIT disrupted the syntrophic interaction between acetogens and methanogens and totally suppressed methanogens' metabolic activities. The VFA production derived from WAS anaerobic fermentation was therefore enhanced due to the interference of antimicrobial MIT stress. This work deciphered dynamic changes and adaptive evolution of anaerobic syntrophic consortia in response to antimicrobial stress and provided guidance on the evaluation and control of the ecological risks of exogenous pollutants in WAS treatment.

Improved Short-Chain Fatty Acids Production and Protein Degradation During the Anaerobic Fermentation of Waste-Activated Sludge via Alumina Slag-Modified Biochar.

As the by-product in the biological sewage treatment, waste-activated sludge (WAS) always suffers from the difficulty of disposal. Anaerobic fermentation to achieve valuable carbon sources is a feasible way for resource utilization of WAS, whereas the process is always restricted by its biochemical efficiency. Hence, the WAS was used as the feedstock in this study. Alumina slag-modified biochar (Al@BioC) respectively from pine wood (PW) or fresh vinegar residue (FVR) was employed to stimulate the process of short-chain fatty acids (SCFAs) production during the anaerobic treatment of WAS. The results indicate that the addition of Al@BioC could facilitate the distinct increase in SCFAs yield (42.66 g/L) by 14.09% and acetate yield (33.30 g/L) by 18.77%, respectively, when compared with that in regular fermentation without Al@BioC addition. Furthermore, protein degradation was also improved. With the Al@BioCPW added, the maximum concentration of soluble protein reached 867.68 mg/L and was 24.39% higher than the initial level, while the enhancement in the group with Al@BioCFVR and without biochar addition was 12.49% and 7.44%, respectively. According to the results of 16S rDNA sequencing, the relative abundance of acid-producing bacteria (Bacteroidota and Firmicutes) was enriched, enhancing the pathways of protein metabolisms and the ability to resist the harsh environment, respectively. Moreover, Proteiniphilum under Bacteroidota and Fastidiosipila under Firmicutes were the main microorganisms to metabolize protein. The above results might provide a novel material for harvesting the SCFAs production, which is conducive to harmless disposal and carbon resource recovery.

Benzethonium chloride affects short chain fatty acids produced from anaerobic fermentation of waste activated sludge: Performance, biodegradation and mechanisms

Benzethonium chloride (BZC) is viewed as a promising disinfectant and widely applied in daily life. While studies related to its effect on waste activated sludge (WAS) anaerobic fermentation (AF) were seldom mentioned before. To understand how BZC affects AF of WAS, production of short chain fatty acids (SCFAs), characteristics of WAS as well as microbial community were evaluated during AF. Results manifested a dose-specific relationship of dosages between BZC and SCFAs and the optimum yield arrived at 2441.01 mg COD/L with the addition of 0.030 g/g TSS BZC. Spectral results and protein secondary structure variation indicated that BZC denatured proteins in the solid phase into smaller proteins or amino acids with unstable structures. It was also found that BZC could stimulate the extracellular polymeric substances secretion and reduce the surface tension of WAS, leading to the enhancement of solubilization. Beside, BZC promoted the hydrolysis stage (increased by 7.09 % to 0.030 g/g TSS BZC), but inhibited acetogenesis and methanogenesis stages (decreased by 6.85 % and 14.75 % to 0.030 g/g TSS BZC). The microbial community was also regulated by BZC to facilitate the enrichment of hydrolytic and acidizing microorganisms (i.e. Firmicutes). All these variations caused by BZC were conducive to the accumulation of SCFAs. The findings contributed to investigating the effect of BZC on AF of WAS and provided a new idea for the future study of AF mechanism.

Disinfectant sodium dichloroisocyanurate synergistically strengthened sludge acidogenic process and pathogens inactivation: Targeted upregulation of functional microorganisms and metabolic traits via self-adaptation

Harmless and resourceful treatment of waste activated sludge (WAS) have been the crucial goal for building environmental-friendly and sustainable society, while the synergistic realization approach is currently limited. This work skillfully utilized the disinfectant sodium dichloroisocyanurate (NaDCC) to simultaneously achieve the pathogenic potential inactivation (decreased by 60.1 %) and efficient volatile fatty acids (VFAs) recovery (increased by 221.9 %) during WAS anaerobic fermentation in rather cost-effective way (Chemicals costs:0.4 USD/kg VFAs versus products benefits: 2.68 USD/kg chemical). Mechanistic analysis revealed that the C=O and NCl bonds in NaDCC could spontaneously absorb sludge (binding energy -4.9 kJ/mol), and then caused the sludge disintegration and organic substrates release for microbial utilization due to the oxidizability of NaDCC. The disruption of sludge structure along with the increase of bioavailable fermentation substrates contributed to the selectively regulation of microbial community via enriching VFAs-forming microorganisms (e.g., Pseudomonas and Streptomyces) and reducing VFAs-consuming microorganisms, especially aceticlastic methanogens (e.g., Methanothrix and Methanospirillum). Correspondingly, the metabolic functions of membrane transport, substrate metabolism, pyruvate metabolism, and fatty acid biosynthesis locating in the central pathway of VFAs production were all upregulated while the methanogenic step was inhibited (especially acetate-type methanogenic pathway). Further exploration unveiled that for those enriched functional anaerobes were capable to activate the self-adaptive systems of DNA replication, SOS response, oxidative stress defense, efflux pump, and energy metabolism to counteract the unfavorable NaDCC stress and maintain high microbial activities for efficient VFAs yields. This study would provide a novel strategy for synergistic realization of harmless and resourceful treatment of WAS, and identify the interrelations between microbial metabolic regulations and adaptive responses.

Surfactant and antibiotic co-occurrence reshaped the acidogenic process for volatile fatty acids production during sludge anaerobic fermentation

The overuse of surfactants and antibiotics has led to their high concentration in waste activated sludge (WAS), and these exogenous pollutants have been shown to pose various influences on the subsequent anaerobic treatment process. Previous works have primarily concerned the impacts of individual pollutants on WAS anaerobic fermentation process. This work revealed the synergetic effects of sodium dodecyl benzene sulfonate (SDBS) and sulfadiazine (SDZ) co-occurrence in WAS on the biosynthesis of volatile fatty acids (VFAs). The addition of SDBS in the SDZ reactor significantly increased VFAs generation, and this increase was correlated with the concentration of SDZ. The VFAs production exhibited a 200.0–211.9 % and 5.9–20.4 % increase in comparison with the sole SDZ and SDBS reactor, respectively. The SDBS and SDZ co-occurrence facilitated the solubilization, hydrolysis, and acidification stages of WAS fermentation synchronously. SDBS was effectively to disintegrate the cemented structure of extracellular polymeric substances and meanwhile improve the SDZ solubilization, which increase the SDZ bioavailability as well as biotoxicity to the anaerobic species. Herein, the anaerobic consortia structure was evidently reshaped, and the keystone microbes Acetoanaerobium and Fususibacter, as well-tolerated hydrolytic-acidogenic bacteria, were greatly enriched. Furthermore, the functional microbial metabolic traits responsible for the substrate extracellular hydrolysis (e.g., glsA and MAN2C1), intracellular metabolism (e.g., ALDO and asdA), and fatty acid generation (e.g., aarC) were all upregulated in the SDBS/SDZ co-occurrence reactor.

Contribution identification of hydrolyzed products of potassium ferrate on promoting short-chain fatty acids production from waste activated sludge

Potassium ferrate (K2FeO4) has been extensively employed to promote short-chain fatty acids (SCFAs) production from anaerobic fermentation of waste activated sludge (WAS) because of its potent oxidizing property and formation of alkaline hydrolyzed products (potassium hydroxide, KOH and ferric hydroxide, Fe(OH)3). However, whether K2FeO4 actually works as dual functions of both an oxidizing agent and an alkalinity enhancer during the anaerobic fermentation process remains uncertain. This study aims to identify the contributions of hydrolyzed products of K2FeO4 on SCFAs production. The results showed that K2FeO4 did not execute dual functions of oxidization and alkalinity in promoting SCFAs production. The accumulation of SCFAs using K2FeO4 treatment (183 mg COD/g volatile suspended solids, VSS) was less than that using either KOH (192 mg COD/g VSS) or KOH & Fe(OH)3 (210 mg COD/g VSS). The mechanism analysis indicated that the synergistic effects caused by oxidization and alkalinity properties of K2FeO4 did not happen on solubilization, hydrolysis, and acidogenesis stages, and the inhibition effect caused by K2FeO4 on methanogenesis stage at the initial phase was more severe than that of its hydrolyzed products. It was also noted that the inhibition effects of K2FeO4 and its hydrolyzed products on the methanogenesis stage could be relieved during a longer sludge retention time, and the final methane yields using KOH or KOH & Fe(OH)3 treatment were higher than that using K2FeO4, further confirming that dual functions of K2FeO4 were not obtained. Therefore, K2FeO4 may not be an alternative strategy for enhancing the production of SCFAs from WAS compared to its alkaline hydrolyzed products. Regarding the strong oxidization property of K2FeO4, more attention could be turned to the fates of refractory organics in the anaerobic fermentation of WAS.

Thiosulfate pretreatment enhancing short-chain fatty acids production from anaerobic fermentation of waste activated sludge: Performance, metabolic activity and microbial community

A novel strategy based on thiosulfate pretreatment for enhancing short-chain fatty acids (SCFAs) from anaerobic fermentation (AF) of waste activated sludge (WAS) was proposed in this study. The results showed that the maximal SCFA yield increased from 206.1 ± 4.7 to 1097.9 ± 17.2 mg COD/L with thiosulfate dosage increasing from 0 to 1000 mg S/L, and sulfur species contribution results revealed that thiosulfate was the leading contributor to improve SCFA yield. Mechanism exploration disclosed that thiosulfate addition largely improved WAS disintegration, due to thiosulfate serving as a cation binder for removing organic-binding cations, especially Ca2+ and Mg2+, dispersing the extracellular polymeric substance (EPS) structure and further entering into the intracellularly by stimulated carrier protein SoxYZ and subsequently caused cell lysis. Typical enzyme activities and related functional gene abundances indicated that both hydrolysis and acidogenesis were remarkably enhanced while methanogenesis was substantially suppressed, which were further strengthened by the enriched hydrolytic bacteria (e.g. C10-SB1A) and acidogenic bacteria (e.g. Aminicenantales) but severely reduced methanogens (e.g. Methanolates and Methanospirillum). Economic analysis confirmed that thiosulfate pretreatment was a cost-effective and efficient strategy. The findings obtained in this work provide a new thought for recovering resource through thiosulfate-assisted WAS AF for sustainable development.

Insights into the Effects of CeO2 Nanoparticles on Medium-Chain Carboxylates Production from Waste Activated Sludge

The synthesis of medium-chain carboxylates (MCCs) from waste-activated sludge (WAS) upgrading has received considerable attention. However, limited research has been conducted on the effects of CeO2 nanoparticles (NPs) on this process. This study showed that 1 mg/g−TS of CeO2 NPs improved the solubilization of WAS, resulting in higher production of MCCs. At 5 mg/g−TS, CeO2 NPs weakly inhibited 3 biological steps. Despite this, there was an enhancement in WAS solubilization, thus the overall production of MCCs was similar to the control. However, doses of CeO2 NPs ranging from 25–100 mg/g−TS were unable to offset biological inhibition, leading to a decrease in MCC production. The toxic mechanisms involved were not the generation of reactive oxygen species or Ce ions from CeO2 NPs to anaerobic sludge, but instead the decline of extracellular polymeric substance (EPS) and destruction of the cell membrane through physical penetration. Microbial community analysis confirmed that 1 mg/g−TS of CeO2 NPs increased the relative abundance of key bacteria involved in the anaerobic fermentation of WAS. The MCC microbe Clostridium sensu stricto was enriched in the control group, while the relative abundance of this genus was significantly reduced with 100 mg/g−TS CeO2 NPs.

Revealing an unrecognized role of free ammonia in sulfur transformation during sludge anaerobic treatment

Free ammonia (FA), the unionized form of ammonium, is presented in anaerobic fermentation of waste activated sludge (WAS) at high levels. However, its potential role in sulfur transformation, especially H2S production, during WAS anaerobic fermentation process was unrecognized previously. This work aims to unveil how FA affects anaerobic sulfur transformation in WAS anaerobic fermentation. It was found that FA significantly inhibited H2S production. With an increase of FA from 0.04 to 159 mg/L, H2S production reduced by 69.9%. FA firstly attacked tyrosine-like proteins and aromatic-like proteins in sludge EPSs, with CO groups being responded first, which decreased the percentage of α-helix/(β-sheet + random coil) and destroyed hydrogen bonding networks. Cell membrane potential and physiological status analysis showed that FA destroyed membrane integrity and increased the ratio of apoptotic and necrotic cells. These destroyed sludge EPSs structure and caused cell lysis, thus strongly inhibited the activities of hydrolytic microorganisms and sulfate reducing bacteria. Microbial analysis showed that FA reduced the abundance of functional microbes (e.g., Desulfobulbus and Desulfovibrio) and genes (e.g., MPST, CysP, and CysN) involved in organic sulfur hydrolysis and inorganic sulfate reduction. These findings unveil an actually existed but previously overlooked contributor to H2S inhibition in WAS anaerobic fermentation.

Surfactant enhances anaerobic fermentative hydrogen sulfide production: Changes in sulfur-containing organics structure and microbial community

The presence of surfactants in waste activated sludge (WAS) system is generally regarded as beneficial to sludge treatment such as enhancing sludge dewatering and improving value-added fermentation products generation. However, in this study, it was firstly found that sodium dodecylbenzene sulfonate (SDBS, a typical surfactant) obviously increased toxic hydrogen sulfide (H2S) gas production from WAS anaerobic fermentation at environmentally relevant concentrations. Experimental results showed that H2S production from WAS significantly increased from 53.24 × 10−3 to 111.25 × 10−3 mg/g volatile suspended solids (VSS) when SDBS level increased from 0 to 30 mg/g total suspended solid (TSS). It was found that SDBS presence destroyed WAS structure and enhanced sulfur containing organics release. SDBS reduced the proportion of α-helix structure, damaged disulfide bridges and protein conformation, and effectively destroyed protein structure. SDBS promoted sulfur containing organics degradation and provided more readily hydrolyzed micro-molecule organics for sulfide production. Microbial analysis showed that SDBS addition enhanced the abundance of functional genes encoding protease, ATP-binding cassette transporters, and amino acids lyase, enhanced the activities and abundance of hydrolytic microbes, thus increased sulfide production from the hydrolysis of sulfur containing organics. Compared with the control, 30 mg/g TSS SDBS increased organic sulfurs hydrolysis and amino acids degradation by 47.1 % and 63.5 %, respectively. Key genes analysis further showed that SDBS addition promoted sulfate transport system and dissimilatory sulfate reduction. SDBS presence also lowered fermentation pH, promoted the chemical equilibrium transformation of sulfide, thus increased H2S gas release.

Identification and degradation of structural extracellular polymeric substances in waste activated sludge via a polygalacturonate-degrading consortium

By maintaining the cell integrity of waste activated sludge (WAS), structural extracellular polymeric substances (St-EPS) resist WAS anaerobic fermentation. This study investigates the occurrence of polygalacturonate in WAS St-EPS by combining chemical and metagenomic analyses that identify ∼22% of the bacteria, including Ferruginibacter and Zoogloea, that are associated with polygalacturonate production using the key enzyme EC 5.1.3.6. A highly active polygalacturonate-degrading consortium (GDC) was enriched and the potential of this GDC for degrading St-EPS and promoting methane production from WAS was investigated. The percentage of St-EPS degradation increased from 47.6% to 85.2% after inoculation with the GDC. Methane production was also increased by up to 2.3 times over a control group, with WAS destruction increasing from 11.5% to 28.4%. Zeta potential and rheological behavior confirmed the positive effect which GDC has on WAS fermentation. The major genus in the GDC was identified as Clostridium (17.1%). Extracellular pectate lyases (EC 4.2.2.2 and 4.2.2.9), excluding polygalacturonase (EC 3.2.1.15), were observed in the metagenome of the GDC and most likely play a core role in St-EPS hydrolysis. Dosing with GDC provides a good biological method for St-EPS degradation and thereby enhances the conversion of WAS to methane.

Surfactant aggravated the antibiotic's stress on antibiotic resistance genes proliferation by altering antibiotic solubilization and microbial traits in sludge anaerobic fermentation

The excessive application of antibiotics and surfactants resulted in their massive accumulation in waste activated sludge (WAS), but the co-occurrent impacts of antibiotics and surfactants on the antibiotic resistant genes (ARGs) fates have seldom reported. This work mainly revealed the roles and critical mechanisms of sodium dodecyl benzene sulfonate (SDBS) on the sulfadiazine (SDZ) stressing for ARGs distribution during WAS anaerobic fermentation. High-throughput qPCR and metagenomic analysis revealed that SDBS aggravated the SDZ selective pressure, and accelerated the proliferation of ARGs. The total abundance of ARGs was increased from 8.81 × 1010 in SDZ to 1.17 × 1011 copies/g TSS in the SDBS/SDZ co-occurrence system. Specifically, the absolute abundances of ermF (MLSB), mefA (MLSB), tetM-01 (tetracycline), tetX (tetracycline), sul2 (sulfonamide) and strB (aminoglycoside) were risen from 4.60 × 108–7.44 × 109 copies/g TSS in the SDZ reactor to 1.02 × 109–4.63 × 1010 copies/g TSS in SDBS/SDZ reactor. SDBS was contributed to the SDZ solubilization and simultaneously effective in disintegrating extracellular polymeric substances and improving cell membrane permeability, which would facilitate the SDZ transport and its interactions with ARGs hosts. Consequently, the microbial community structure was evidently altered, and the typical ARGs hosts (i.e., Saccharimonadales and Ahniella) were greatly enriched. Also, the cell signal transduction systems (i.e., glnL, glrK and pilG), oxidative stress response (i.e., frmA and recA) and bacterial secretion systems (i.e., VirB4), which were related with ARGs propagation, were all provoked in the co-occurred SDBS/SDZ reactor compared with that of sole SDZ. PLS-PM analysis suggested that the bacterial community was the predominant factor that determined the ARGs fates, followed by mobile genetic elements and metabolic pathway. This work demonstrated the interactive effects of surfactants and antibiotics on the ARGs fates in WAS fermentation systems and gave insightful implications on the ecological risks of different exogenous pollutants.

Sulfite pretreatment enhances the medium-chain fatty acids production from waste activated sludge anaerobic fermentation

Production of high-value medium chain fatty acids (MCFAs) from anaerobic fermentation of waste activated sludge (WAS) has been considered as a promising alternative for renewable energy resources. However, the low biodegradability of WAS greatly limits the anaerobic fermentation performance. This study proposed and demonstrated a novel approach, sulfite pretreatment, to efficiently produce MCFAs through anaerobic fermentation of WAS. Pretreatment of WAS at a sulfite concentration of 100–500 mg S/L for 24 h effectively improved the MCFAs production and MCFAs selectivity and the promotion effect was positively correlated with the sulfite concentration used in pretreatment (Pearson's R > 0.9). The maximum MCFAs production of 6.84 g COD/L and MCFAs selectivity of 39.1 % were both achieved under 500 mg S/L sulfite pretreatment, which accounts for 2.6 times and 2.4 times of the control, respectively (MCFAs production of 2.62 g COD/L and MCFAs selectivity of 16.4 % in the control). Sulfite pretreatment also enhanced the WAS degradation from 25 ± 2 % in the control to a maximum of 39 ± 2 % under 500 mg S/L sulfite pretreatment. The electron transfer efficiency and COD flows from the substrate to products were enhanced by up to 25 % due to the sulfite pretreatment, which supports the enhanced WAS degradation. Sulfite pretreatment also promoted the solubilization, hydrolysis, and acidification processes during the anaerobic fermentation by up to 200 %, 60 %, and 45 %, respectively, which subsequently makes more substrates available for MCFAs production. The findings from this study provide a potential solution of using industrial sulfite-laden wastes for WAS pretreatment, to enhance the MCFAs production at a minimized cost.

Effects of free nitrous acid combined with alkyl polyglucoside on short-chain fatty acids production from waste activated sludge anaerobic fermentation and fermentation liquor for polyhydroxyalkanoates synthesis

Free nitrous acid (FNA) pretreatment and alkyl polyglucoside (APG) addition are environmentally-friendly methods for enhancing waste activated sludge (WAS) anaerobic fermentation (AF) performances. The short-chain fatty acids (SCFAs) produced are available substrates for polyhydroxyalkanoates (PHAs) synthesis to reduce the C-source cost. This study aims to investigate the optimal fermentation conditions of FNA + APG treatment and the performance of PHAs synthesis by the SCFAs from AF system. Through investigating the FNA + APG combined treatment effects, key enzyme activities, and microbial community variation, the AFL from optimal fermentation condition was utilized as a C-source for PHAs synthesis. Experimental results indicated that 0.653 mg/L FNA + 0.075 g/g VSS APG treatment could synergistically achieve the maximum total SCFAs yield of 324.94 mg COD/g VSS at day 5 of AF, which was much more than those of sole FNA or APG treatment. Mechanism analysis indicated that FNA + APG treatment could promote WAS solubilization, hydrolysis and acidogenesis, while severely inhibiting methanogenesis. Microbial community analysis results demonstrated that SCFA producers, including Enterococcus and Clostridium, became the dominant genera in the FNA + APG reactor. Finally, AF liquor after ammonia removal was applied for batch-mode PHAs synthesis, and PHA yields increased to a maximum of 44.44 % of biomass (w/w) after five operation periods.

Nonmonotonic effect of CuO nanoparticles on medium-chain carboxylates production from waste activated sludge

The growing applications of CuO nanoparticles (NPs) in industrial and agriculture has increased their concentrations in wastewater and subsequently accumulated in waste activated sludge (WAS), raising concerns about their impact on reutilization of WAS, especially on the medium-chain carboxylates (MCCs) production from anaerobic fermentation of WAS. Here we showed that CuO NPs at 10–50 mg/g-TS can significantly inhibit MCCs production, and reactive oxygen species generation was revealed to be the key factor linked to the phenomena. At lower CuO NPs concentrations (0.5–2.5 mg/g-TS), however, MCCs production was enhanced, with a maximum level of 37% compared to the control. The combination of molecular approaches and metaproteomic analysis revealed that although low dosage CuO NPs (2.5 mg/g-TS) weakly inhibited chain elongation process, they displayed contributive characteristics both in WAS solubilization and transport/metabolism of carbohydrate. These results demonstrated that the complex microbial processes for MCCs production in the anaerobic fermentation of WAS can be affected by CuO NPs in a dosage-dependent manner via regulating microbial protein expression level. Our findings can provide new insights into the influence of CuO NPs on anaerobic fermentation process and shed light on the treatment option for the resource utilization of CuO NPs polluted WAS.

Insights into the effect of carbamazepine on the anaerobic fermentation of waste activated sludge: Performance, mechanisms and regulation

The effects of pollutants on anaerobic fermentation of waste activated sludge (WAS) have been widely studied. Nevertheless, little attention was paid to how to mitigate the adverse effects of pollutants on anaerobic fermentation. In this work, representative pollutant carbamazepine (CBZ) was chosen to explore the potential effects of CBZ on WAS anaerobic fermentation. Experimental results showed that CBZ at environmental levels slightly promoted the production of short-chain fatty acids (SCFAs), which increased from 3858.2 to 4143.8 mg COD/L as the CBZ dose increased from 8 to 28 mg/kg VSS, but further increasing the CBZ dose to 88 mg/kg VSS decreased the production of SCFAs. Context of homologous group database revealed that the abundance of some functional traits changed, such as RNA processing and modification, energy production and conversion, and amino acid transport. This adverse effect was sustained because CBZ could not be degraded during anaerobic fermentation. Therefore, a regulation strategy was proposed, e.g., persulfate was used to attenuate the toxicity of CBZ to WAS anaerobic fermentation, and the results showed that the removal of CBZ reached 83.7 % in the persulfate-treated reactor. Based on density functional theory and metabolic intermediates determination, hydroxylation and deamination reactions were the two main metabolic pathways for CBZ degradation.

The fate of diclofenac in anaerobic fermentation of waste activated sludge

Diclofenac (DCF), a nonsteroidal anti-inflammatory drug, is one of the most commonly detected pharmaceuticals in wastewater treatment plants. However, the fate of DCF in waste activated sludge (WAS) anaerobic fermentation has not been well-understood so far. This work therefore aims to comprehensively reveal whether and how DCF is transformed in WAS mesophilic anaerobic fermentation through both experimental investigation and density functional theory (DFT) calculation. Experimental results showed that ∼28.8% and 45.8% of DCF were respectively degraded during the batch and long-term fermentation processes. Based on the detected intermediates and DFT-predicted active sites, three metabolic pathways, i.e., chlorination, hydroxylation, and dichlorination, responsible for DCF transformation were proposed. DFT calculation also showed that the Gibbs free energy (ΔG) of the three transformation pathways was respectively 19.0, −4.3, and −19.3 kcal/mol, suggesting that the latter two reactions (i.e., hydroxylation and dichlorination) were thermodynamically favorable. Illumina MiSeq sequencing analyses revealed that DCF improved the populations of complex organic degradation microbes such as Proteiniclasticum and Tissierellales, which was in accord with the chemical analyses above. This work updates the fundamental understanding of the degradation of DCF in WAS anaerobic fermentation process and enlightens engineers to apply theoretical calculation to the field of sludge treatment or other complex microbial ecosystems.