Phages in sludge from the A/O wastewater treatment process play an important role in the transmission of ARGs.

Phages can influence the horizontal gene transfer (HGT) of antibiotic resistance genes (ARGs) through transduction, but their profiles and effects on the transmission of ARGs are unclear, especially in complex swine sludge. In this study, we investigated the characterization of phage and ARG profiles in sludge generated from anoxic/oxic (A/O) wastewater treatment processes on swine farms using metagenomes and viromes. The results demonstrated that 205-221 subtypes of ARGs could be identified in swine sludge, among which sul1, tet(M), and floR were the dominant ARGs, indicating that sludge is an important reservoir of ARGs, especially in sludge (S) tanks. The greater abundance of mobile genetic elements (MGEs) in the S tank could significantly contribute to the greater abundance of ARGs there compared to the anoxic (A) and oxic (O) tanks (P < 0.05). However, when we compared the abundances of ARGs and MGEs in the A and O tanks, we observed opposite significant differences (P < 0.05), suggesting that MGEs are not the only factor influencing the abundance of ARGs. The high proportion of lysogenic phages in sludge from the S tank can also have a major impact on the ARG profile. Siphoviridae, Myoviridae, and Podoviridae were the dominant phage families in sludge, and a network diagram of bacteria-ARG-phages revealed that dominant phages and bacteria acted simultaneously as potential hosts for ARGs, which may have led to phage-mediated HGT of ARGs. Therefore, the risk of phage-mediated HGT of ARGs cannot be overlooked.

Comparing methanol and glycerol as carbon sources for mainstream partial denitrification/anammox in an IFAS process.

This study explored the implementation of mainstream partial denitrification with anammox (PdNA) in the second anoxic zone of a wastewater treatment process in an integrated fixed film activated sludge (IFAS) configuration. A pilot study was conducted to compare the use of methanol and glycerol as external carbon sources for an IFAS PdNA startup, with a goal to optimize nitrogen removal while minimizing carbon usage. The study also investigated the establishment of anammox bacteria on virgin carriers in IFAS reactors without the use of seeding, and it is the first IFAS PdNA startup to use methanol as an external carbon source. The establishment of anammox bacteria was confirmed in both reactors 102 days after startup. Although the glycerol-fed reactor achieved a higher steady-state maximum ammonia removal rate because of anammox bacteria (1.6 ± 0.3 g/m2/day) in comparison with the methanol-fed reactor (1.2 ± 0.2 g/m2/day), both the glycerol- and methanol-fed reactors achieved similar average in situ ammonia removal rates of 0.39 ± 0.2 g/m2/day and 0.40 ± 0.2 g/m2/day, respectively. Additionally, when the upstream ammonia versus NOx (AvN) control system maintained an ideal ratio of 0.40-0.50 g/g, the methanol-fed reactor attained a lower average effluent TIN concentration (3.50 ± 1.2 mg/L) than the glycerol-fed reactor (4.43 ± 1.6 mg/L), which was prone to elevated nitrite concentrations in the effluent. Overall, this research highlights the potential for PdNA in IFAS configurations as an efficient and cost-saving method for wastewater treatment, with methanol as a viable carbon source for the establishment of anammox bacteria. PRACTITIONER POINTS: Methanol is an effective external carbon source for an anammox startup that avoids the need for costly alternative carbon sources. The methanol-fed reactor demonstrated higher TIN removal compared with the glycerol-fed reactor because of less overproduction of nitrite. Anammox bacteria was established in an IFAS reactor without seeding and used internally stored carbon to reduce external carbon addition. Controlling the influent ammonia versus NOx (AvN) ratio between 0.40 and 0.50 g/g allowed for low and stable TIN effluent conditions.

Description and Genomic Characteristics of Diaphorobacter limosus sp. nov., Isolated from a Sewage-Treatment Plant.

A Gram-stain-negative, rod-shaped, non-motile, catalase-positive, denitrifying bacterium, designated strain Y-1T, was isolated from an aeration tank of a sewage treatment plant in China and characterized using polyphasic taxonomic approaches. Strain Y-1T could grow at 10-37 °C (optimum 25 °C), at pH 5.0-10.0 (optimum 7.0) and in the presence of 0-3.0% (w/v) NaCl (optimum 0.5%). The phylogenetic tree based on the 16S rRNA gene sequences revealed that strain Y-1T was a member of genus Diaphorobacter, and showed the highest sequence similarities with Diaphorobacter oryzae RF3T (97.50%), Diaphorobacter nitroreducens NA10BT (97.38%) and Diaphorobacter aerolatus 8604S-37T (96.56%). In terms of carbon source utilization and enzyme activities, strain Y-1T was significantly different from its similar strains. The major respiratory quinone was Q-8, and the main polar lipid was phosphatidylethanolamine. Comparative genomic analysis of strain Y-1T and other Diaphorobacter species was conducted to explore the mechanisms underlying the differences among these strains. Strain Y-1T encoded 3957 genes, consisting of 3813 protein-coding genes and 144 RNA coding genes, and encoded 652 enzymes with 31 unique enzymes compared with other related species. The DNA G + C content was 69.95 mol%. Strain Y-1T exhibited 41.71% DNA-DNA relatedness and 95% ANIb with the most related type strains.On the basis of the evidence presented from polyphasic analysis, strain Y-1T was suggested as a novel species within the genus Diaphorobacter, for which the name Diaphorobacter limosus sp. nov. is proposed, with the type strain Y-1T (= KCTC 92852T = CCTCC AB 2023032T).

Unveiling the influence of microaeration and sludge recirculation on enhancement of pharmaceutical removal and microbial community change of the novel anaerobic baffled biofilm - membrane bioreactor in treating building wastewater.

This research aims to conduct a comparative investigation of the role played by microaeration and sludge recirculation in the novel anaerobic baffled biofilm-membrane bioreactor (AnBB-MBR) for enhancing pharmaceutical removal from building wastewater. Three AnBB-MBRs - R1: AnBB-MBR, R2: AnBB-MBR with microaeration and R3: AnBB-MBR with microaeration and sludge recirculation - were operated simultaneously to remove Ciprofloxacin (CIP), Caffeine (CAF), Sulfamethoxazole (SMX) and Diclofenac (DCF) from real building wastewater at the hydraulic retention time (HRT) of 30 h for 115 days. From the removal profiles of the targeted pharmaceuticals in the AnBB-MBRs, it was found that the fixed-film compartment (C1) could significantly reduce the targeted pharmaceuticals. The remaining pharmaceuticals were further removed with the microaeration compartment. R2 exhibited the utmost removal efficiency for CIP (78.0 %) and DCF (40.8 %), while SMX was removed most successfully by R3 (microaeration with sludge recirculation) at 91.3 %, followed by microaeration in R2 (88.5 %). For CAF, it was easily removed by all AnBB-MBR systems (>90 %). The removal mechanisms indicate that the microaeration in R2 facilitated the adsorption of CIP onto microaerobic biomass, while the enhanced biodegradation of CAF, SMX and DCF was confirmed by batch biotransformation kinetics and the adsorption isotherms of the targeted pharmaceuticals. The microbial groups involved in biodegradation of the targeted compounds under microaeration were identified as nitrogen removal microbials (Nitrosomonas, Nitrospira, Thiobacillus, and Denitratisoma) and methanotrophs (Methylosarcina, Methylocaldum, and Methylocystis). Overall, explication of the integration of AnBB-MBR with microaeration (R2) confirmed it as a prospective technology for pharmaceutical removal from building wastewater due to its energy-efficient approach characterized by minimal aeration supply.

Oleispirillum naphthae gen. nov., sp. nov., a bacterium isolated from oil sludge, and proposal of Oleispirillaceae fam. nov.

A microaerophilic, Gram-negative, motile, and spiral-shaped bacterium, designated Y-M2T, was isolated from oil sludge of Shengli oil field. The optimal growth condition of strain Y-M2T was at 25 °C, pH 7.0, and in the absence of NaCl. The major polar lipid was phosphatidylethanolamine. The main cellular fatty acid was iso-C17  :  0 3-OH. It contained Q-9 and Q-10 as the predominant quinones. The DNA G+C content was 68.1 mol%. Strain Y-M2T showed the highest 16S rRNA gene sequence similarity to Telmatospirillum siberiense 26-4bT (91.1 %). Phylogenetic analyses based on 16S rRNA gene and genomes showed that strain Y-M2T formed a distinct cluster in the order Rhodospirillales. Genomic analysis showed that Y-M2T possesses a complete nitrogen-fixation cluster which is phylogenetically close to that of methanogene. The nif cluster, encompassing the nitrogenase genes, was found in every N2-fixing strain within the order Rhodospirillales. Phylogeny, phenotype, chemotaxonomy, and genomic results demonstrated that strain Y-M2T represents a novel species of a novel genus in a novel family Oleispirillaceae fam. nov. in the order Rhodospirillales, for which the name Oleispirillum naphthae gen. nov., sp. nov. was proposed. The type strain is Y-M2T (=CCAM 827T=JCM 34765T).

New insights into functional divergence and adaptive evolution of uncultured bacteria in anammox community by complete genome-centric analysis.

Anaerobic ammonium-oxidation (anammox) bacteria play a crucial role in global nitrogen cycling and wastewater nitrogen removal, but they share symbiotic relationships with various other microorganisms. Functional divergence and adaptive evolution of uncultured bacteria in anammox community remain underexplored. Although shotgun metagenomics based on short reads has been widely used in anammox research, metagenome-assembled genomes (MAGs) are often discontinuous and highly contaminated, which limits in-depth analyses of anammox communities. Here, for the first time, we performed Pacific Biosciences high-fidelity (HiFi) long-read sequencing on the anammox granule sludge sample from a lab-scale bioreactor, and obtained 30 accurate and complete metagenome-assembled genomes (cMAGs). These cMAGs were obtained by selecting high-quality circular contigs from initial assemblies of long reads generated by HiFi sequencing, eliminating the need for Illumina short reads, binning, and reassembly. One new anammox species affiliated with Candidatus Jettenia and three species affiliated with novel families were found in this anammox community. cMAG-centric analysis revealed functional divergence in general and nitrogen metabolism among the anammox community members, and they might adopt a cross-feeding strategy in organic matter, cofactors, and vitamins. Furthermore, we identified 63 mobile genetic elements (MGEs) and 50 putative horizontal gene transfer (HGT) events within these cMAGs. The results suggest that HGT events and MGEs related to phage and integration or excision, particularly transposons containing tnpA in anammox bacteria, might play important roles in the adaptive evolution of this anammox community. The cMAGs generated in the present study could be used to establish of a comprehensive database for anammox bacteria and associated microorganisms. These findings highlight the advantages of HiFi sequencing for the studies of complex mixed cultures and advance the understanding of anammox communities.

Temperature-phased anaerobic sludge digestion effectively removes antibiotic resistance genes in a full-scale wastewater treatment plant.

Sludge is a major by-product and the final reservoir of antibiotic resistance genes (ARGs) in wastewater treatment plants (WWTPs). Temperature-phased anaerobic digestion (TPAD), consisting of thermophilic anaerobic digestion (AD) (55 °C) and mesophilic AD processes (37 °C), has been implemented in WWTPs for sludge reduction while improving the biomethane production. However, the impact of TPAD on the ARGs' fate is still undiscovered in lab-scale experiments and full-scale WWTPs. This study, for the first time, investigated the fate of ARGs during the TPAD process across three seasons in a full-size WWTP. Ten typical ARGs and one integrase gene of class 1 integron (intI1) involving ARGs horizontal gene transfer were examined in sludge before and after each step of the TPAD process. TPAD reduced aac(6')-Ib-cr, blaTEM, drfA1, sul1, sul2, ermb, mefA, tetA, tetB and tetX by 87.3-100.0 %. TPAD reduced the overall average absolute abundance of targeted ARGs and intI1 by 92.39 % and 92.50 %, respectively. The abundance of targeted ARGs in sludge was higher in winter than in summer and autumn before and after TPAD. During the TPAD processes, thermophilic AD played a major role in the removal of ARGs, contributing to >60 % removal of ARGs, while the subsequent mesophilic AD contributed to a further 31 % removal of ARGs. The microbial community analysis revealed that thermophilic AD reduced the absolute abundance of ARGs hosts, antibiotic resistant bacteria. In addition, thermophilic AD reduced the abundance of the intI1, while the intI1 did not reproduce during the mesophilic AD, also contributing to a decline in the absolute abundance of ARGs in TPAD. This study demonstrates that TPAD can effectively reduce the abundance of ARGs in sludge, which will suppress the transmission of ARGs from sludge into the natural environment and deliver environmental and health benefits to our society.

Study on the mechanism of action of methane production by co-fermentation of sludge and lignite.

To improve the methanogenic efficiency of lignite anaerobic fermentation and explore innovative approaches to sludge utilization, a co-fermentation technique involving lignite and sludge was employed for converting biomass into biomethane. Volatile suspended solids were introduced as a native enrichment of the sludge and mixed with lignite for fermentation. The synergistic fermentation mechanism between sludge and lignite for biomethane production was analyzed through biochemical methane potential experiments, measurement of various parameters pre- and post-fermentation, observation of bacterial population changes during the peak of reaction, carbon migration assessment, and evaluation of rheological characteristics. The results showed that the addition of sludge in the anaerobic fermentation process improved the microorganisms' ability to degrade lignite and bolstered biomethane production. Notably, the maximum methane production recorded was 215.52 mL/g-volatile suspended solids, achieved at a sludge to coal ratio of 3:1, with a synergistic growth rate of 25.37%. Furthermore, the removal rates of total suspended solids, and total chemical oxygen demand exhibited an upward trend with an increasing percentage of sludge in the mixture. The relative abundance and activity of the methanogens population were found to increase with an appropriate ratio of sludge to lignite. This observation confirmed the migration of carbon between the solid-liquid-gas phases, promoting enhanced system affinity. Additionally, the changes in solid-liquid phase parameters before and after the reaction indicated that the addition of sludge improved the system's degradation capacity. The results of the study hold significant implications in realizing the resource utilization of sludge and lignite while contributing to environmental protection endeavors.

Metabolic Profiles and Microbial Synergy Mechanism of Anammox Biomass Enrichment and Membrane Fouling Alleviation in the Anammox Dynamic Membrane Bioreactor.

The anammox dynamic membrane bioreactor (DMBR) is promising in applications with enhanced anammox biomass enrichment and fouling alleviation. However, the metabolic mechanism underlying the functional features of anammox sludge and the biofilm membrane is still obscure. We investigated the metabolic networks of anammox sludge and membrane biofilm in the DMBR. The cooperation between anammox and dissimilatory nitrate reduction to ammonium processes favored the robust anammox process in the DMBR. The rapid bacterial growth occurred in the DMBR sludge with 1.33 times higher biomass yield compared to the MBR sludge, linked to the higher activities of lipid metabolism, nucleotide metabolism, and B vitamin-related metabolism of the DMBR sludge. The metabolism of the DMBR biofilm microbial community benefited the fouling alleviation that the abundant fermentative bacteria and their cooperation with the anammox sludge microbial community promoted organics degradation. The intensified degradation of foulants by the DMBR biofilm community was further evidenced by the active carbohydrate metabolism and the upregulated vitamin B intermediates in the biofilms of the DMBR. Our findings provide insights into key metabolic mechanisms for enhanced biomass enrichment and fouling control of the anammox DMBR, guiding manipulations and applications for overcoming anammox biomass loss in the treatment of wastewater under detrimental environmental conditions.

Novel three-sludge municipal wastewater treatment process coupling denitrifying phosphorus removal with anaerobic ammonium oxidation.

The two-sludge anoxic dephosphation (DEPHANOX) process frequently encounters the challenge of elevated effluent ammonia levels in practical applications. In this study, the anaerobic ammonium oxidation (anammox) biofilm was introduced into the DEPHANOX system, transforming it into a three-sludge system, enabling synchronous nitrogen and phosphorus elimination, particularly targeting ammonia. Despite a chemical oxygen demand/total nitrogen ratio of 4.3 ± 0.8 in the actual municipal wastewater and 4.5 h of aeration, the effluent total nitrogen was 13.7 mg/L, lower than the parallel wastewater treatment plant. Additionally, the effluent ammonia reduced to 5.1 ± 2.5 mg/L. Notably, denitrifying phosphorus removal and anammox were coupled in the anoxic zone, yielding 74.5 % nitrogen and 87.8 % phosphorus removal. 16S rRNA gene sequencing identified denitrifying phosphorus-accumulating organisms primarily in floc sludge (Saprospiraceae 7.07 %, Anaerolineaceae 1.95 %, Tetrasphaera 1.57 %), while anammox bacteria inhabited the biofilm (Candidatus Brocadia 4.00 %). This study presents a novel process for efficiently treating municipal wastewater.

Anaerobic digestion of wastewater from hydrothermal liquefaction of sewage sludge and combined wheat straw-manure.

Hydrothermal liquefaction (HTL) shows promise for converting wet biomass waste into biofuel, but the resulting high-strength process water (PW) requires treatment. This study explored enhancing energy recovery by anaerobic digestion using semi-batch reactors. Co-digesting manure with HTL-PW from wheat straw-manure co-HTL yielded methane (43-49% of the chemical oxygen demand, COD) at concentrations up to 17.8 gCOD·L-1, whereas HTL-PW from sewage sludge yielded methane (43% of the COD) up to only 12.8 gCOD·L-1 and complete inhibition occurred at 17 gCOD·L-1. Microbial community shifts confirmed inhibition of methanogenic archaea, while hydrolytic-fermentative bacteria were resilient. Differences in chemical composition, particularly higher levels of N-containing heterocyclic compounds in PW of sewage sludge, likely caused the microbial inhibition. The considerable potential of combining HTL with anaerobic digestion for enhanced energy recovery from straw-manure in an agricultural context is demonstrated, yet sewage sludge HTL-PW requires more advanced approaches to deal with methanogenesis inhibitors.

Enhancing wastewater treatment efficiency: A hybrid technology perspective with energy-saving strategies.

The study aimed to investigate how hybrid technology, combined with various intermittent aeration (IA) strategies, contributes to reducing the energy costs of wastewater treatment while simultaneously ensuring a high treatment efficiency. Even with IA subphases lasting half as long as those without aeration, and oxygen levels reduced from 3.5 to 1.5 mg O2/L, pollutants removal efficiency remains robust, allowing for a 1.41-fold reduction in energy consumption (EO). Hybrid technology led to a 1.34-fold decrease in EO, along with improved denitrification efficiency from 74.05 ± 4.71 to 81.87 ± 2.43 % and enhanced biological phosphorus removal from 35.03 ± 4.25 to 87.32 ± 3.64 %. The high nitrification efficiency may have been attributed to the abundance of Pseudomonas, Acinetobacter, and Rhodococcus, which outcompeted the genera of autotrophic nitrifying bacteria, suggesting that the hybrid system is favorable for the growth of heterotrophic nitrifiers.

Advanced nitrogen and phosphorus removal in pilot-scale anaerobic/aerobic/anoxic system for municipal wastewater in Northern China.

Removing nitrogen and phosphorus from low ratio of chemical oxygen demand to total nitrogen and temperature municipal wastewater stays a challenge. In this study, a pilot-scale anaerobic/aerobic/anoxic sequencing batch reactor (A/O/A-SBR) system first treated 15 m3/d actual municipal wastewater at 8.1-26.4 °C for 224 days. At the temperature of 15.7 °C, total nitrogen in influent and effluent were 45.5 and 10.9 mg/L, and phosphorus in influent and effluent were 3.9 and 0.1 mg/L. 16 s RNA sequencing results showed the relative abundance of Competibacter and Tetrasphaera raised to 1.25 % and 1.52 %. The strategy of excessive, no and normal sludge discharge enriched and balanced the functional bacteria, achieving an endogenous denitrification ratio more than 43.3 %. Sludge reduction and short aerobic time were beneficial to energy saving contrast with a Beijing municipal wastewater treatment. This study has significant implications for the practical application of the AOA-SBR process.

Towards zero excess sludge discharge with built-in ozonation for wastewater biological treatment.

In this study, a biological treatment process, which used a built-in ozonation bypass to achieve sludge reduction, was built to treat the industrial antifreeze production wastewater (mainly composed of ethylene glycol). The results indicated there is a positive correlation between ozone dosage and sludge reduction. At the laboratory level, the MLSS in the system can be stably controlled at around 3400 mg MLSS L-1 under the dosage of 0.18 g O3 g-1 MLSS. Ozonation can increase the compactness of sludge flocs (fractal dimension increased from 1.89 to 1.92). Ozone destroys microbial cell membranes and alters the structure of sludge flocs through direct oxidation through electrophilic reactions. It leads to the release of intracellular polysaccharides, proteins, and other biological macromolecules in microorganisms, thereby promoting the implicit growth of microbial populations. Some bacteria such as g_Pseudomonas, g_Gemmobacter, etc. have strong ethylene glycol degradation ability and tolerance to ozonation. The removal of ethylene glycol includes the glyoxylate cycle, glycine serine carbon cycle, and the glutamate-cysteine ligase pathway of assimilation. Gene KatG and gpx may be key factors in improving microbial tolerance to ozonation. The comprehensive evaluation from the perspectives of cost and carbon emission shows that choosing ozone cracking-implicit growth in wastewater treatment systems has significant cost advantages and application value.

Evaluation of various carbon sources on ammonium assimilation and denitrifying phosphorus removal in a modified anaerobic-anoxic-oxic process from low-strength wastewater.

A pilot-scale continuous-flow modified anaerobic-anoxic-oxic (MAAO) process examined the impact of external carbon sources (acetate, glucose, acetate/propionate) on ammonium assimilation, denitrifying phosphorus removal (DPR), and microbial community. Acetate exhibited superior efficacy in promoting the combined process of ammonia assimilation and DPR, enhancing both to 50.0 % and 60.0 %, respectively. Proteobacteria and Bacteroidota facilitated ammonium assimilation, while denitrifying phosphorus-accumulating organisms (DPAOs) played a key role in nitrogen (N) and phosphorus (P) removal. Denitrifying glycogen-accumulating organisms (DGAOs) aided N removal in the anoxic zone, ensuring stable N and P removal and recovery. Acetate/propionate significantly enhanced DPR (77.7 %) and endogenous denitrification (37.9 %). Glucose favored heterotrophic denitrification (29.6 %) but had minimal impact on ammonium assimilation. These findings provide valuable insights for wastewater treatment plants (WWTPs) seeking efficient N and P removal and recovery from low-strength wastewater.

Competitive bio-augmentation overcoming unusual direct inhibitor inefficacy in mainstream nitrite-oxidizing bacteria suppression: Unveiling the underpinnings in microbial and nitrogen metabolism aspects.

The long-stabilized mainstream partial nitritation/Anammox (PN/A) process continues to encounter significant challenges from nitrite-oxidizing bacteria (NOB). Therefore, this study aimed to determine an efficient, rapid, and easily implementable strategy for inhibiting NOB. A laboratory-scale reactor was operated continuously for 325 days, experiencing NOB outbreak in mainstream and recovery with simulated sidestream support. The results show that direct inhibitory strategies including intermittent aeration and approximately 35 mg/L free ammonia had unusual weak inhibitory effects on NOB activity. Subsequently, the exogenous Anammox from sidestream employed as a competitive bio-augmentation approach rapidly inhibited NOB dynamics. Evidence suggests that the damaged hydroxyapatite granules under low pH conditions might have contributed to NOB dominance by diminishing Anammox bacteria activity, thereby creating a substrate-rich environment favoring NOB survival. In contrast, the introduction of exogenous Candidatus Kuenenia facilitated the nitrogen removal efficiency from 32.5 % to over 80 %. This coincided with a decrease in the relative abundance of Nitrospira from 16.5 % to 2.7 % and NOB activity from 0.34 to 0.07 g N/(g mixed liquor volatile suspended solid)/d. Metagenomic analysis reveals a decrease in the functional potential of most nitrite transport proteins, coupled with a significant increase in eukaryotic-like serine/threonine-protein kinase involved in cellular regulation, during the Anammox activity recovery. This study's findings reveal the feasibility of the bio-augmentation based on substrate competition, wherein sidestream processes support the mainstream PN/A integration, offering significant potential for practical applications.

Enhancing anammox process with granular activated carbon: A study on Microbial Extracellular Secretions (MESs).

Granular activated carbon (GAC), a porous carbon-based material, provides increased attachment space for functional microorganisms and enhances nitrogen removal by facilitating extracellular electron transfer in the anammox process. This study investigates the effects of GAC on the biosynthesis of microbial extracellular secretions (MESs) and explores the roles of these secretions in anammox activities. Four lab-scale reactors were operated: two downstream UASB reactors (D1 and D2) receiving effluents from the upstream UASB reactors (U1: no-GAC, U2: yes-GAC). Our results indicate that MESs were enhanced with the addition of GAC. The effluent from U2 exhibited a 59.62 % higher amino acid content than that from U1. These secretions contributed to an increase in the nitrogen loading rate (NLR) in the downstream reactors. Specifically, NLR in D1 increased from 130.5 to 142.7 g N/m3/day, and in D2, it escalated from 137.5 to 202.8 g N/m3/day, likely through acting as cross-feeding substrates or vital nutrients. D2 also showed increased anammox bacterial activity, enriched Ca. Brocadia population and hao gene abundance. Furthermore, this study revealed that D2 sludge has significantly higher extracellular polymeric substances (EPS) (48.71 mg/g VSS) and a larger average granule size (1.201 ± 0.119 mm) compared to D1 sludge. Overall, GAC-stimulated MESs may have contributed to the enhanced performance of the anammox process.

Redox mediator chlorophyll accelerates low-temperature biological denitrification with responses of extracellular polymers and changes in microbial community composition.

Low temperatures limit the denitrification wastewater in activated sludge systems, but this can be mitigated by addition of redox mediators (RMs). Here, the effects of chlorophyll (Chl), 1,2-naphthoquinone-4-sulfonic acid (NQS), humic acid (HA), and riboflavin (RF), each tested at three concentrations, were compared for denitrification performance at low temperature, by monitoring the produced extracellular polymeric substances (EPS), and characterizing microbial communities and their metabolic potential. Chl increased the denitrification rate most, namely 4.12-fold compared to the control, followed by NQS (2.62-fold increase) and HA (1.35-fold increase), but RF had an inhibitory effect. Chl promoted the secretion of tryptophan-like and tyrosine-like proteins in the EPS and aided the conversion of protein from tightly bound EPS into loosely bound EPS, which improved the material transfer efficiency. NQS, HA, and RF also altered the EPS components. The four RMs affected the microbial community structure, whereby both conditionally abundant taxa (CAT) and conditionally rare or abundant taxa (CRAT) were key taxa. Among them, CRAT members interacted most with the other taxa. Chl promoted Flavobacterium enrichment in low-temperature activated sludge systems. In addition, Chl promoted the abundance of nitrate reduction genes narGHI and napAB and of nitrite reduction genes nirKS, norBC, and nosZ. Moreover, Chl increased abundance of genes involved in acetate metabolism and in the TCA cycle, thereby improving carbon source utilization. This study increases our understanding of the enhancement of low-temperature activated sludge by RMs, and demonstrates positive effects, in particular by Chl.

Influence of adding strategy of biochar on medium-chain fatty acids production from sewage sludge.

Production of medium-chain fatty acids (MCFAs) from sewage sludge has dual effects on valuable sludge disposal and renewable energy generation, while low efficiency limits its application. Biochar addition is considered an effective method to improve MCFAs production. In this study, the influence of biochar adding strategies (i.e., adding biochar in acidification or chain elongation (CE) processes) on MCFAs production was explored. Results showed that by adding biochar in the acidification process, MCFAs accumulation increased by over 114%, accompanied by the highest carbon conversion efficiency (134.66%) and electron transfer efficiency of MCFAs (94.22%) by the terminal CE. Adding biochar before the acidification process better enriched CE bacteria (e.g., Paraclostridium) and strengthened the dominant metabolic pathway. In contrast, the biochar added before the CE process priorly enriched the bacteria capable of degrading organics, like unclassified_f__Dysgonomonadaceae, norank_f__norank_o__OPB41, and Acetobacterium. The differences in excessive ethanol oxidation and short-chain fatty acids accumulation induced by varied adding strategies might be responsible for this.

Research on carbon and nitrogen removal of tetramethylammonium hydroxide containing wastewater by combined anaerobic/integrated fixed film activated sludge process.

Tetramethylammonium hydroxide (TMAH) is widely used as a developer and etchant in the thin-film transistor liquid crystal display industry, which is the main component of developer wastewater with low C/N ratio. This study investigated TMAH degradation by combined anaerobic/integrated fixed film activated sludge (A/IFAS) process, especially for nitrogen removal. Effects of process condition on the TMAH degradation were studied, including dissolved oxygen concentration in IFAS reactor and the temperature of anaerobic reactor. Especially, the nitrogen removal was studied through the monitoring of intermediate products during TMAH biodegradation. The results indicated that lower the anaerobic treatment temperature can provide more available organic matters to enhance the denitrification in the subsequent IFAS reactor. Less oxygen supply in the IFAS reactor contributed to simultaneous nitrification and denitrification. Removal efficiency of total organic carbon and total nitrogen was up to 95.8% and 80.7%, when the temperature of anaerobic treatment was controlled at 30 °C with the DO kept at 0.7 mg/L. It indicated that A/IFAS process was efficient in carbon and nitrogen removal for TMAH degradation. The results also confirmed intermediate products of TMAH biodegradation can be used as the electron donor during denitrification, including trimethylamine, dimethylamine and methylamine. Illumina MiSeq sequencing showed that Proteobacteria was the dominant phylum contribute to nitrogen removal. Compared to sludge flocs in IFAS reactor, richer community and higher microbial diversity were observed in the biofilm.