Influence of C/N ratio and ammonia on nitrogen removal and N2O emissions from one-stage partial denitrification coupled with anammox.

The development of low carbon treatment processes is an important issue worldwide. Partial denitrification coupled with anammox (PD/A) is a novel strategy to remove nitrogen and reduce N2O emissions. The influence of C/N ratio and NH4+ concentration on nitrogen removal and N2O emissions was investigated in batch reactors filled with PD/A coupled sludge. A C/N ratio of 2.1 was effective for nitrogen removal and N2O reduction; higher ammonia concentration might make anammox more active and indirectly reduce N2O emissions. Long-term operation further confirmed that a C/N ratio of 2.1 resulted in a minimum effluent N2O concentration (mean value of 0.94 µmol L-1); as the influent NH4+ concentration decreased to 50 mg L-1 (NH4+-N/NO3--N: 1), the nitrogen removal rate increased to 82.41%. Microbial analysis showed that anammox bacteria (Candidatus Jettenia and Ca. Brocadia) were enriched in the PD/A system and Ca. Brocadia gradually dominated the anammox community, with the relative abundance increasing from 1.69% to 18.44% between days 97 and 141. Finally, functional gene analysis indicated that the abundance of nirS/K and hao involved in partial denitrification and anammox, respectively, increased during long-term operation of the reactor; this change benefitted nitrogen metabolism in anammox, which could indirectly reduce N2O emissions.

Effects of intrathecally administered interferon α on chronic constriction injury model rats' mechanical pain threshold and G protein expression in the spinal cord.

INTRODUCTION: The aim of the study was to explore the analgesic mechanism of effects of intrathecally administered interferon a (IFN-a) on chronic constriction injury (CCI) model rats. MATERIAL AND METHODS: 24 rats were divided into 6 groups, with 4 rats in each group, including the negative control group (Group N, no operation or treatment), the sham operation group (Group S, only the left sciatic nerve of the rats was exposed without ligation, 0.9% NaCl was intrathecally administered), and four experimental groups (CCI model was established first and then different drugs were intrathecally administered respectively), including 0.9% NaCl (Group C), IFN-a (Group CI), morphine (Group CM), and IFN-a combined with morphine (Group CIM). The mRNA levels of G proteins in both the spinal cord and dorsal root ganglia (DRG), as well as the content of amino acid and chemokine (C-X-C motif) ligand 6 (CXCL-6) in the cerebrospinal fluid were measured and analysed in each group. RESULTS: Intrathecal administration of IFN-a increased the mechanical pain threshold in CCI rats (33.32 ±1.36 vs. 21.08 ±1.59, p < 0.001), achieving the effect comparable to that of morphine (33.32 ±1.36 vs. 32.44 ±3.18, p > 0.05), increased the mRNA expression level of Gi protein (0.62 ±0.04 vs. 0.49 ±0.05, p = 0.006), and decreased the mRNA expression level of Gs protein in the spinal cord (1.80 ±0.16 vs. 2.06 ±0.15, p = 0.035) and DRG (2.11 ±0.10 vs. 2.79 ±0.13, p < 0.001). The intrathecal administration of both IFN-a and morphine can reduce the glutamate content in the cerebrospinal fluid (261.55 ±38.12 vs. 347.70 ±40.69, p = 0.012), but without any statistically significant difference in the content of CXCL-6 across all groups ( p > 0.05). CONCLUSIONS: Intrathecal injection of IFN-a improved the mechanical pain threshold in CCI rats, so we inferred that intrathecal administration of IFN-a had analgesic effects on neuropathic pain, possibly related to the activation of G-proteincoupled µ receptors in the spinal cord and the inhibition of glutamate release.

Insight into the spatiotemporal distribution of antibiotic resistance genes in estuarine sediments during long-term ecological restoration.

In this study, we aimed to investigate the long-term spatiotemporal changes in hydrodynamics, antibiotics, nine typical subtypes of antibiotic resistance genes (ARGs), class 1 integron gene (intI1), and microbial communities in the sediments of a semi-enclosed estuary during ecological restoration with four treatment stages (influent (#1), effluent of the biological treatment area (#2), oxic area (#3), and plant treatment area (#4)). Ecological restoration of the estuary reduced common pollutants (nitrogen and phosphorus) in the water, whereas variations in ARGs showed noticeable seasonal and spatial features. The absolute abundance of ARGs at sampling site #2 considerably increased in autumn and winter, while it significantly increased at sampling site #3 in spring and summer. The strong intervention of biological treatment (from #1 to #2) and aerators (from #2 to #3) in the estuary substantially affected the distribution of ARGs and dominant antibiotic-resistant bacteria (ARB). The dominant ARB (Thiobacillus) in estuarine sediments may have low abundance but important dissemination roles. Meanwhile, redundancy and network analysis revealed that the microbial communities and intl1 were key factors related to ARG dissemination, which was affected by spatial and seasonal ecological restoration. A positive correlation between low flow velocity and certain ARGs (tetM, tetW, tetA, sul2, and ermC) was observed, implying that flow optimization should also be considered in future ecological restoration to remediate ARGs. Furthermore, the absolute abundance of ARGs can be utilized as an index to evaluate the removal capacity of ARGs by estuarine restoration.

Metagenomic insights into the spatiotemporal responses of antibiotic resistance genes and microbial communities in aquaculture sediments.

The dissemination of antibiotic resistance genes (ARGs) in aquaculture systems is a potential threat to environmental safety and human health. However, the spatiotemporal distribution pattern of ARGs and key factors associated with their dissemination in aquaculture sediments remain unclear. In this study, ARGs, mobile genetic elements, microbial community composition, heavy metal contents, and nutrient contents of samples collected from a whole culture cycle of fish in a representative aquaculture farm were characterized. The distribution patterns of nine subtypes of ARGs (tetW, tetM, tetA, ermC, ermB, sul1, sul2, floR, and qnrS) showed clear spatiotemporal differences. The absolute abundance of ARGs in aquaculture sediments was higher in winter and in rivers of the aquaculture farm. Proteobacteria was the dominant phylum in all sediment samples. The results of network and redundancy analyses confirmed that the Dechloromonas, Candidatus Accumulibacter, Smithella, Geobacter, and Anaeromyxobacter belonging to Proteobacteria were positively correlated with ARGs, suggesting that these microbial species are potential hosts of corresponding ARGs. Our study highlights that the microbial community is the determining factor for ARG dissemination. Strategies for inhibiting these potential hosts of ARGs should be developed based on controllable factors.

Integrated moving bed biofilm reactor with partial denitrification-anammox for promoted nitrogen removal: Layered biofilm structure formation and symbiotic functional microbes.

Partial denitrification/anaerobic ammonia oxidation (anammox) (PD/A) is currently an advanced nitrogen removal process. This study developed a PD/A system in a moving bed biofilm reactor. Results showed that the nitrogen removal efficiency reached 76.60% with a COD/NO3-N of 2.0, and the contribution of anammox was 88.01%. Further analysis showed that the biocarriers could form layered pH and dissolved oxygen structures to promote the aggregation of different functional bacteria at various depths, thus stabilizing the coupled process. Microbial structure analysis showed that the abundance of Saccharimonadales, responsible for denitrification, increased from 0% to 36.27% between day 0 and day 120, while the abundance of Candidatus Jettenia, responsible for anammox, decreased from 10.41% to 2.20%. The synergistic effect of Saccharimonadales and Candidatus Jettenia enabled stable and efficient removal of nitrogen. This study proposed a novel configuration of the PD/A process and provided a theoretical basis for its promotion and application.

The response of sediment microbial communities to temporal and site-specific variations of pollution in interconnected aquaculture pond and ditch systems.

Sediment microbial communities play critical roles in the health of fish and the biogeochemical cycling of elements in aquaculture ecosystems. However, the response of microbial communities to temporal and spatial variations in interconnected aquaculture pond and ditch systems remains unclear. In this study, 61 sediment bacterial samples were collected over one year from 11 sites (including five ponds and six ditches) in a 30-year-old fish aquaculture farm. The 16S rRNA approach was used to determine the relative abundances of microbial communities in the sediment samples. The relationships among nutrients, heavy metals, and abundant microorganisms were analyzed. Our results showed that Proteobacteria, Bacteroides and Chloroflexi were the predominant phyla in the sediments of aquaculture pond, with average abundances of 36.33%, 18.60%, and 14.58%, respectively. The microbial diversity in aquaculture sediments was negatively correlated (P < 0.05) with the concentrations of total nitrogen and total phosphorus in sediments, indicating that the microbial diversity is highly associated with the remediation of nutrients in sediments. The sediment samples with high similarities were discovered by the t-distributed stochastic neighbor embedding (t-SNE) method. The site-specific correlations between specific microorganisms and heavy metals were explored. The network analysis revealed that the microbial diversities in aquaculture ponds were more stable than that in aquaculture ditches. The network analysis also illustrated that the microbial genera with low relative abundances may become key groups of microbial communities in sediment ecosystems. Our work deepens the understanding of the relationships between microbial communities and the spatiotemporal characteristics of surface water and sediments in aquaculture farms.

Altered Neural Processing of Reward and Punishment in Women With Methamphetamine Use Disorder.

It has been suggested that the altered function of reward and punishment is an important vulnerability factor leading to the development of drug use disorders. Previous studies have identified evidence of neurophysiological dysfunction in the reward process of individuals with substance use disorders. To date, only a few event-related potential (ERP) studies have examined the neural basis of reward and punishment processing in women with methamphetamine (MA) use disorders. The current ERP research aims to investigate the neurophysiological mechanisms of reward and punishment in women with MA use disorder using a monetary incentive delay task. Nineteen women with MA use disorder (MA group) and 20 healthy controls (HC group) were recruited in this study. The behavioral data showed that the reaction time (RT) was faster and the response accuracy (ACC) was higher for the potential reward and punishment conditions compared to neutral conditions. During the monetary incentive anticipation stage, the Cue-P3, and stimulus-preceding negativity (SPN) were larger in the MA group than in the HC group. The SPN under the potential reward condition was larger than that under the neutral condition in the MA group but not in the HC group. During the monetary incentive consummation stage, the feedback-related negativity and feedback P3 (FB-P3) following positive feedback were significantly larger than negative feedback in the potential reward condition for the HC group, but not for the MA group. However, the FB-P3 following negative feedback was significantly larger than positive feedback in the potential punishment condition for the MA group, but not the HC group. The results suggest that women with MUD have stronger expectations of generic reward and stronger response of generic harm avoidance, which could be targeted in designing interventions for women with MA use disorder.

Facilitating biofilm formation of Pseudomonas aeruginosa via exogenous N-Acy-L-homoserine lactones stimulation: Regulation on the bacterial motility, adhesive ability and metabolic activity.

The N-Acy-L-homoserine lactones (AHLs) mediated quorum sensing (QS) system exhibited important ecological significance in bacterial biofilm formation. However, the previous studies mainly focused on indigenous AHLs while the role of exogenous AHLs has remained unclear. This study evaluated the roles of exogenous AHLs on the biofilm formation of Pseudomonas aeruginosa. Both the C6-HSL and C8-HSL promoted the biofilm formation of P. aeruginosa with an enhancement of 2.47 and 1.88 times, respectively. Further analysis showed that exogenous AHLs contributed greatly to the adhesive ability instead of growth rate. Also, the bacterial motility and metabolic activities were significantly improved by AHLs. Moreover, the microbial functional genes (i.e. lasI, lasR, rhlI and rhlR) involved in regulating the biofilm formation were highly expressed in AHLs reactors. These findings expanded the knowledge of AHLs functions in mediating biofilm formation, and provided insightful guidance on the biofilm regulation in the wastewater treatment via biofilm technology.

Enhancement of the photosynthetic and removal performance for microalgae-based technologies by co-culture strategy and strigolactone induction.

In this study, digestate and biogas were treated simultaneously based on fungi-microalgae symbionts and fungi-microalgae-bacteria symbionts, respectively. The effects of strigolactone (GR24) on the biomass of symbionts and photosynthesis of algae, and purification of digestate and biogas were explored. With the induction of GR24, the metabolism of microorganism-microalgae systems and photosynthesis of microalgae were enhanced causing the rapid growth of the symbionts, further strengthening the purification performance of the systems. Moreover, the GR24 enhanced the CO2 removal performance of the co-culture systems via increasing the carbonic anhydrase activity of microalgae cells. The fungi-microalgae-bacteria co-culture strategy with GR24 induction was found to obtain great photosynthetic and removal performance, possessing the 73.54 ± 6.83% CO2 removal rate and 88.61 ± 8.24% COD removal rate.

Co-culturing microalgae with endophytic bacteria increases nutrient removal efficiency for biogas purification.

Endophytic bacteria were isolated from Chlorella vulgaris and co-cultured with its host microalgae to determine whether this symbiotic system is suitable for purifying biogas and biogas slurry. Results showed that endophytic bacteria S395-1 and S395-2 belonged to different genera. Both strains promoted microalgae growth while improving photosynthetic performance, carbonic anhydrase activity, nutrient removal efficiency, and CO2 fixation. The optimal bacteria (S395-2)-to-microalgae ratio and co-culture duration were 10:1 and 7 days. Under this condition, the growth rate and carbonic anhydrase activity were 0.196 ± 0.06 d-1 and 31.24 ± 0.28 EU/cell, respectively. The symbiotic system had removal efficiencies of 88.29 ± 5.03%, 88.31 ± 4.29%, 88.21 ± 4.51%, and 68.13 ± 1.69% for chemical oxygen demand, nitrogen, phosphorus, and CO2, respectively. These results will provide a framework for constructing a microalgal-bacterial consortium that can improve wastewater treatment and enhance biogas quality.

Continuous waste activated sludge and food waste co-fermentation for synchronously recovering vivianite and volatile fatty acids at different sludge retention times: Performance and microbial response.

A practical approach of synchronously recovering vivianite and volatile fatty acids (VFAs) by food waste (FW) and waste activated sludge (WAS) co-fermentation in continuous operation was investigated. Approximately 82.88% P as high-purity vivianite (95.23%) and 7894 mg COD/L VFAs were finally recovered. The simultaneous addition of FW and FeCl3 contributed to the fermentation conditions by adjusting pH biologically and increasing the concentration of organic substrates, which enhanced the Fe3+ reduction efficiency and microbial activities (e.g., hydrolases and acidogenic enzymes). Microbial analysis found the functional bacteria related to Fe3+ reduction and VFAs generation were further enhanced and enriched. Besides, results indicated that the efficiencies of Fe2+ and P release and VFAs recovery were highly linked to SRT, the satisfactory fermentation performance was obtained at SRT of 6 d. This research would provide a practical waste recycling technology to treat FW and WAS simultaneously for recovering vivianite and VFAs synchronously.

Effect of GR24 concentrations on biogas upgrade and nutrient removal by microalgae-based technology.

Freshwater microalgae Chlorella vulgaris was cultured and induced with strigolactone (GR24) to simultaneously eliminate nutrients in biogas slurry and purify biogas. Treatment with 10-7 M GR24 yielded maximum growth rate and mean daily productivity for algae at 0.187 ± 0.06 d-1 and 0.097 ± 0.008 g L-1 d-1, respectively. Results from chlorophyll fluorescence transients method demonstrated that moderate concentration of GR24 could enhance the photosynthetic performance of microalgae. In addition, GR24 affected intracellular carbonic anhydrase activity and chlorophyll-a content. Maximum chemical oxygen demand, total nitrogen, and CO2 removal efficiencies were 78.62 ± 2.36%, 76.47 ± 1.53% and 64.05 ± 1.15% with 10-7 M GR24 induction, respectively. Further, highest total phosphorus removal efficiency (80.27 ± 1.93%) was observed at 10-9 M. The optimal GR24 concentration range was determined to be between 10-9 and 10-7 M in consideration with nutrient and CO2 removal efficiencies.

Social Incentives Anticipation and Consummation: Investigating Neural Activity in Women Using Methamphetamine.

Individuals with methamphetamine use disorder are considered to have enhanced reactivity to rewards or drug cues. However, whether this holds true in the social incentives processing is still unclear. The current study investigated the electroencephalographical (EEG) evidence of social incentives processing in women with methamphetamine use disorder (MA group, n = 19) and in a healthy control group (HC group, n = 20) using social incentive delay (SID) tasks. In the SID, participants received a "Like" (e.g., thumbs up) or "Unlike" (e.g., thumbs down) from WeChat emojis as social incentives, or neutral feedback. During the anticipation stage, the Cue-P3 and stimulus-preceding negativity (SPN) were larger for the social incentives condition than for the neutral condition. During the consummation stage, the feedback-related negativity (FRN) was marginally significantly larger in the HC group than the MA group for the social incentive condition, whereas there was no significant difference between the groups for neutral condition. Interestingly, the FB-P3 was larger for social positive feedback than for social negative feedback in the MA group, but not in HC group. Furthermore, only the HC group showed significant positive correlation between the anticipatory event-related brain potential (ERP, such as Cue-P3) and the consummatory ERP (FB-P3) in the social incentive condition. The findings suggest that women with MA use disorder have a blunted neural response to the processing of social incentives and a blunted neural response to negative social feedback, which helps to elucidate the neural mechanisms of social incentives processing in individuals with MA use disorder.

Revealing hydrodynamic effects on flocculation performance and surface properties of sludge by comparing aeration and stirring systems via computational fluid dynamics aided calculation.

The effects of aeration and stirring systems on the physical properties of sludge were analyzed using a computational fluid dynamics (CFD) model. The aims of this study were to (1) compare the effects of aeration and stirring on sludge properties using the same turbulent mixing intensity, and (2) to reveal the relationship between sludge properties and hydrodynamic indicators to determine how hydrodynamic conditions influence sludge flocculation. Mixing experiments with stirring and aeration were carried out in 2-L beakers with the average velocity gradient (G) set to 90, 190, or 280 s-1. The sludge flocculation performance, zeta potential, and Gibbs free energy (ΔG) were analyzed and the flow velocity, turbulence energy, turbulence dissipation rate, and Kolmogorov microscale were calculated as hydrodynamic parameters. The average flow velocity and the turbulence dissipation rate were obviously higher in the stirring system than in the aeration system at the same G. However, the turbulence energy and Kolmogorov microscale in the aeration system were much higher than those in the stirring system. Both the zeta potential and ΔG were lower in the aeration system than the stirring system. The zeta potential and ΔG results for the two systems suggest that aeration is more beneficial for sludge flocculation than stirring even though the sludge flocculation performance F/F0 in the stirring and aeration systems showed no obvious differences. Significant relationships between hydrodynamic parameters calculated based on the CFD model and average values of sludge properties in the stable phase showed that the Kolmogorov microscale, average flow velocity, and turbulence energy were appropriate hydrodynamic parameters for evaluating the flocculation performance F/F0, zeta potential, and ΔG, respectively.

Physicochemical and Biological Effects on Activated Sludge Performance and Activity Recovery of Damaged Sludge by Exposure to CeO2 Nanoparticles in Sequencing Batch Reactors.

Recently, the growing release of CeO2 nanoparticles (CeO2 NPs) into sewage systems has attracted great concern. Several studies have extensively explored CeO2 NPs' potential adverse impacts on wastewater treatment plants; however, the impaired activated sludge recovery potentials have seldom been addressed to date. To explore the physicochemical and biological effects on the activated sludge performance and activity recovery of damaged sludge by exposure to CeO2 NPs in sequencing batch reactors (SBRs), four reactors and multiple indicators including water quality, key enzymes, microbial metabolites, the microbial community structure and toxicity were used. Results showed that 10-week exposure to higher CeO2 NP concentration (1, 10 mg/L) resulted in a sharp decrease in nitrogen and phosphorus removal efficiencies, which were consistent with the tendencies of key enzymes. Meanwhile, CeO2 NPs at concentrations of 0.1, 1, and 10 mg/L decreased the secretion of tightly bound extracellular polymeric substances to 0.13%, 3.14%, and 28.60%, respectively, compared to the control. In addition, two-week recovery period assays revealed that the functional bacteria Proteobacteria, Nitrospirae and Planctomycetes recovered slightly at the phyla level, as analyzed through high-throughput sequencing, which was consistent with the small amount of improvement of the effluent performance of the system. This reflected the small possibility of the activity recovery of damaged sludge.

Phosphorus recovery as vivianite from waste activated sludge via optimizing iron source and pH value during anaerobic fermentation.

This study presented an innovative method for phosphorus (P) recovery as vivianite from waste activated sludge (WAS) via optimizing iron dosing and pH value during anaerobic fermentation (AF). The optimal conditions for vivianite formation were in the pH range of 6.0-9.0 with initial PO43- >5 mg/L and Fe/P molar ratio of 1.5. Notably, FeCl3 showed advantages over ZVI for the simultaneous release of Fe2+ and PO43- during WAS fermentation, especially in acidic conditions. The FeCl3 dosing at pH 3.0 could contribute to 78.81% Fe2+ release and 85.69% of total PO43- release from WAS. They were ultimately recovered in the form of high-purity vivianite (93.67%). Clostridiaceae (40.25%) was the predominant bacteria in FeCl3-pH3 reactors, which played key roles in inducing dissimilatory iron reduction for Fe2+ formation. Therefore, P recovery as vivianite from WAS fermentation might be a promising and highly valuable approach to relieve the P crisis.

Review on the determination and distribution patterns of a widespread contaminant artificial sweetener in the environment.

The accurate determination of widespread artificial sweeteners (ASs) and the information of their distributions in environments are of significance to investigate the environmental behaviors. This paper firstly reviews the typical analytic methodologies for ASs and the main influencing factors during the analytic processes. Solid-phase extraction (SPE) with LC-ESI-MS is currently the leading-edge method. However, the efficiency and accuracy for ASs analysis in environmental samples are also dependent on the SPE cartridges, buffers and pH, matrix effects, and sample stability. A basic procedure for ASs determination in different environmental samples is proposed. The current occurrences of ASs in environments are then evaluated. The ASs, especially the acesulfame and sucralose, are widely detected in various environmental medium. The concentrations of investigated ASs are generally in the order of wastewater treatment plants (WWTPs) influent > WWTPs effluent > surface water > groundwater > drinking water; and atmosphere > soil. The ASs levels in the environment exhibit significant differences among different regions. Further analysis indicates that the phenomenon is highly correlated with the consumption patterns and the removal efficiency of WWTPs in a specific country.

Potentials and challenges of phosphorus recovery as vivianite from wastewater: A review.

Due to the shortage of phosphorus resources and the limitations of existing phosphorus recovery methods, phosphorus recovery in the form of vivianite has attracted considerable attention with its natural ubiquity, easy accessibility and foreseeable economic value. This review systematically summarizes the chemistry of vivianite, including the characteristics, formation process and influencing factors of the material. Additionally, the potential of phosphorus recovery as vivianite from wastewater has also been comprehensively examined from the prospects of economic value and engineering feasibility. In general, this method is theoretically and practically feasible, and brings some extra benefits in WWTPs. However, the insufficient understanding on vivianite recovery in wastewater/sludge decelerate the development and exploration of such advanced approach. Further researches and cross-field supports would facilitate the improvement of this technique in the future.

Promoting the anaerobic production of short-chain fatty acids from food wastes driven by the reuse of linear alkylbenzene sulphonates-enriched laundry wastewater.

An efficient attempt to improve the anaerobic fermentation of food wastes (FW) via the reuse of linear alkylbenzene sulphonates (LAS)-enriched laundry wastewater was reported. The production of short-chain fatty acids (SCFAs) from FW was enhanced by approximate 6-folds with high proportions of butyric and valeric acids at appropriate LAS level. Mechanism investigations demonstrated that the solubilization of macromolecule organics in FW was effectively improved by surface tension reduction. The hydrolysis and acidification processes during FW fermentation were accelerated and enhanced with the stimulation of microbial activities (higher activities of hydrolases and ATP concentrations). Also, the abundances of anaerobic microorganisms responsible for SCFAs production were enriched. Metatranscriptomic analysis indicated that the encoding genes involved in the metabolism of fermentation substrates for SCFAs production were highly expressed in LAS-added reactors. It was the simultaneous increase of bioavailable substrates and metabolic activities that contribute to the efficient SCFAs production in LAS-added reactors.

Enhancing the anaerobic bioconversion of complex organics in food wastes for volatile fatty acids production by zero-valent iron and persulfate stimulation.

The addition of zero-valent iron (ZVI) and ZVI & persulfate (PS) were efficient approaches to enhance the production of volatile fatty acids (VFAs), especially butyric acid, from food wastes (FW) during anaerobic fermentation. The maximal concentration of VFAs was increased from 1256 mg COD/L in the control reactor to 8245 mg COD/L with ZVI addition, and it was further improved to 9800 mg COD/L with the PS/ZVI treatment. An investigation of the mechanisms revealed that both the ZVI and PS/ZVI treatments improved the bioavailable substrates in FW and enhanced the bioconversion efficiency of fermentation substrates, especially proteins and lipids. The provision of a sufficient amount of bioavailable substrates was advantageous to the enrichment of the functional bacteria that are responsible for the production of VFAs. Additionally, the microbial activity and key metabolic enzymes involved in the biological VFAs generation processes were stimulated in the ZVI and PS/ZVI-added reactors, which jointly contributed to high-rate VFAs yields.