Temporal Dynamics and Contribution of Phage Community to the Prevalence of Antibiotic Resistance Genes in a Full-Scale Sludge Anaerobic Digestion Plant.

Anaerobic digestion (AD) is an important biological resource recovery process, where microorganisms play key roles for material transformation. There has been some knowledge about the prokaryotic community and antibiotic resistance genes (ARGs) in AD, but there has been very limited knowledge of phages. In this study, samples from a full-scale AD plant were collected over 13 months, sequenced, and analyzed for viral and prokaryotic metagenomes. Totally, 3015 viral operational taxonomic units (vOTUs) were detected, mostly assigned to Caudoviricetes. The phage community had faster temporal variation than the prokaryotic community. Warm seasons harbored a higher abundance of both temperate phages and broad host-range phages. Seven ARGs of 6 subtypes were carried by 20 vOTUs, a representative ermT gene was synthesized and expressed, and the resistance activity in the host was examined, confirming the real activity of virus-carried ARGs in the AD process. Some of the ARGs were horizontally transferred between the phage and prokaryotic genomes. However, phage infection was not found to contribute to ARG transfer. This study provided an insight into the ecological patterns of the phage community, confirmed the antibiotic resistance activity of virus-carried ARGs, evaluated the contribution of phages on the ARG prevalence, and laid the foundation for the control strategies of the community and antibiotic resistance in the AD process.

Enhancing waste sludge solubilization through radio frequency treatment perforating bacterial cells.

Sludge solubilization is known as a rate-limiting step of anaerobic digestion. Although radio frequency (RF) has been applied for sludge pretreatment due to its similar thermal effect as microwave, the potential non-thermal effects of RF treatment remain controversial. In this study, we demonstrate that RF pretreatment enhances the solubilization and lysis of sludge by 8.02%-19.69% through both thermal and non-thermal mechanisms with less energy input. Scanning electron microscope images provide direct evidence that RF-induced microcurrents penetrated bacterial cells, leading to the release of intracellular substances through formed pores. Additionally, the non-thermal effect of RF treatment which could weaken the cell protection and accelerate the lysis rate involves the disruption of binding forces between extracellular polymeric substances and microbial cells. On average, the utilization of RF at a frequency of 27.12 MHz demonstrates its efficacy as a sludge pretreatment technique, as evidenced by a 13.39% reduction in energy consumption and a 16.9% improvement in treatment performance compared to conductive heating (CH). The findings of this study elucidate the possible mechanism of RF treatment of sludge and could establish a theoretical basis for the practical application of RF treatment in sludge management.

Ecological and functional differences of abundant and rare sub-communities in wastewater treatment plants across China.

The microbial community in activated sludge is composed of a small number of abundant sub-community with high abundance and a large number of rare sub-community with limited abundance. Our knowledge regarding the ecological properties of both abundant and rare sub-communities in activated sludge is limited. This article presented an analysis of functional prediction, assembly mechanisms, and biogeographic distribution characteristics of abundant and rare sub-communities in 211 activated sludge samples from 60 wastewater treatment plants across China. Moreover, this study investigated the dominant factors influencing the community structure of these two microbial groups. The results showed that the functions associated with carbon and nitrogen cycling were primarily detected in abundant sub-community, while rare sub-community were primarily involved in sulfur cycling. Both microbial groups were mainly influenced by dispersal limitation, which, to some extent, resulted in a distance-decay relationship in their biogeographic distribution. Moreover, a higher spatial turnover rate of rare sub-communities (0.0887) suggested that spatial differences in microbial community structure among different WWTPs may mainly result from rare sub-community. Moreover, SEM showed that geographic locations affected rare sub-communities greatly, which agreed with their higher dispersal limitation and turnover rate. In contrast, influent characteristics showed stronger correlations with abundant sub-communities, suggesting that abundant sub-community may contribute more to the removal of pollutants. This study enhanced our understanding of abundant and rare microorganisms in activated sludge especially the role of rare species and provided scientific evidence for precise regulation and control of wastewater treatment plants.

Enhancement of medium-chain fatty acids production from sludge anaerobic fermentation liquid under moderate sulfate reduction.

In recent years, there has been a marked increase in the production of excess sludge. Chain-elongation (CE) fermentation presents a promising approach for carbon resource recovery from sludge, enabling the transformation of carbon into medium-chain fatty acids (MCFAs). However, the impact of sulfate, commonly presents in sludge, on the CE process remains largely unexplored. In this study, batch tests for CE process of sludge anaerobic fermentation liquid (SAFL) under different SCOD/SO42- ratios were performed. The moderate sulfate reduction under the optimum SCOD/SO42- of 20:1 enhanced the n-caproate production, giving the maximum n-caproate concentration, selectivity and production rate of 5.49 g COD/L, 21.4% and 4.87 g COD/L/d, respectively. The excessive sulfate reduction under SCOD/SO42- ≤ 5 completely inhibited the CE process, resulting in almost no n-caproate generation. The variations in n-caproate production under different conditions of SCOD/SO42- were all well fitted with the modified Gompertz kinetic model. Alcaligenes and Ruminococcaceae_UCG-014 were the dominant genus-level biomarkers under moderate sulfate reduction (SCOD/SO42- = 20), which enhanced the n-caproate production by increasing the generation of acetyl-CoA and the hydrolysis of difficult biodegradable substances in SAFL. The findings presented in this work elucidate a strategy and provide a theoretical framework for the further enhancement of MCFAs production from excess sludge.

The role of the core microorganisms in the microbial interactions in activated sludge.

In order to gain a deeper understanding of the microbial interactions in wastewater treatment plants (WWTPs) in China and clarify the role of the core community in the microbial interactions in activated sludge (AS), this study used a molecular ecological network approach based on random matrix theory to construct co-occurrence networks of the core microorganisms (CoreN), the whole AS community (WholeN) and the microbial communities without the core microorganisms (OtherN), respectively. It was shown that the WholeN had more complex and tighter connections compared with the OtherN, because of its higher total number of nodes, higher average clustering coefficient, and shorter average geodesic distance. The proportions of positive links in the CoreN, WholeN and OtherN were gradually decreased, indicating that the core microorganisms promoted cooperation between AS microorganisms. Moreover, higher robustness after random removal of 50% of the nodes of the WholeN (0.2836 ± 0.0311) was observed than the robustness of the OtherN (0.1152 ± 0.0263). In addition, the vulnerability of OtherN (0.0514) is significantly higher than WholeN (0.0225). Meanwhile, the average ratio of negative/positive cohesion, was significantly decreased when the core microorganisms were removed. These results demonstrated that core community could strengthen the stability of the ecological network in AS. By discerning the key factors affecting ecological network, AS temperature was observed to have a strong correlation with all three networks. Moreover, pollutants in wastewater shown stronger correlations with the CoreN and WholeN, supporting the point that core community play a critical role in pollutant removal in WWTPs to a certain extent.

Drivers of microbial beta-diversity in wastewater treatment plants in China.

As one of the most well-documented biogeographic patterns, the distance-decay relationship provides insights into the underlying mechanisms driving biodiversity distribution. Although wastewater treatment plants (WWTPs) are well-controlled engineered ecosystems, this pattern has been seen among microbial communities in activated sludge (AS). However, little is known about the relative importance of environmental heterogeneity and dispersal limitation in shaping AS microbial community across China; especially they are related to spatial scale and organism types. Here, we assessed the distance-decay relationship based on different spatial scales and microbial phylogenetic groups by analyzing 132 activated sludge (AS) samples across China comprising 3,379,200 16S rRNA sequences. Our results indicated that the drivers of distance-decay pattern in China were scale-dependent. Microbial biogeographic patterns in WWTPs were mainly driven by dispersal limitation at both local and national scales. In contrast, conductivity, SRT, and pH played dominant roles in shaping AS microbial community compositions at the regional scale. Turnover rates and the drivers of beta-diversity also varied with microorganism populations. Moreover, a quantitative relationship between dispersal limitation ratio and AS microbial turnover rate was generated. Collectively, these results highlighted the importance of considering multiple spatial scales and micro-organism types for understanding microbial biogeography in WWTPs and provided new insights into predicting variations in AS community structure in response to environmental disturbance.

Biogeography and Assembly of Microbial Communities in Wastewater Treatment Plants in China.

Scientific understanding of microbial biogeography and assembly is lacking for activated sludge microbial communities, even though the diversity of microbial communities in wastewater treatment plants (WWTPs) is thought to have a direct influence on system performance. Here, utilizing large-scale 16S rRNA gene data generated from 211 activated sludge samples collected from 15 cities across China, we show activated sludge microbes, whose growth and metabolism can be regulated followed with the metabolic theory of ecology with an apparent Ea value (apparent activation energy) of 0.08 eV. WWTPs at a lower latitude tend to harbor a more diverse array of microorganisms. In agreement with the general understanding, the activated sludge microbial assembly was mainly driven by deterministic processes and the mean annual temperature was identified as the most important factor affecting the microbial community structure. The treatment process types with similar microbial growth types and functions had a distinct impact on the activated sludge microbial community structure only when WWTPs were located near each other and received similar influent. Overall, these findings provide us with a deeper understanding of activated sludge microbial communities from an ecological perspective. Moreover, these findings suggest that, for a given set of performance characteristics (e.g., combined nitrification, denitrification, and phosphorus removal), it may be difficult to employ common engineering levers to control additional aspects of community structure due to the influence of natural environmental factors.

Adsorption and recovery of phosphate from water by amine fiber, effects of co-existing ions and column filtration.

A weak-base adsorption fiber, acrylic amine fiber (AAF), was prepared for removal and recovery of phosphate from water. The adsorption properties of the AAF for phosphate and effects of co-existing ions were investigated using batch and column filtration experiments, scanning electron microscope, and Fourier transform infrared techniques. Experimental results showed that AAF had a high phosphate adsorption capacity of 119 mg/g at pH 7.0. The effects of calcium, sulfate, carbonate, nitrate, and fluoride showed that sulfate and calcium inhibited phosphate adsorption. However, AAF showed higher binding affinity toward phosphate than sulfate. Column filtration results showed that AAF could filter 1420 bed volumes of tap water containing 1.0 mg-P/L of phosphate. The saturated AAF could be regenerated using 0.5 mol/L hydrochloric acid solution and reused. After desorption, phosphate was recovered through precipitation of hydroxyapatite (Ca5(PO4)3OH). The easy of regeneration, good adsorption performance, and the fiber morphology of AAF make it an attractive alternative for phosphate recovery from multiple water sources.

Influence of polyaluminum chloride on microbial characteristics in anaerobic membrane bioreactors for sludge digestion.

In this study, two parallel lab-scale anaerobic membrane bioreactors (AnMBRs), one of which was dosed with polyaluminum chloride (PAC) for membrane fouling control, were operated for treating excess activated sludge collected from a wastewater treatment plant (WWTP). The AnMBRs were inoculated with anaerobic digested sludge collected from an anaerobic digester of another WWTP. The microbial community of digested sludge and cake layer in AnMBRs, as well as that of excess sludge, was analyzed through polymerase chain reaction coupled with denaturing gradient gel electrophoresis (PCR-DGGE) and Illumina MiSeq. The dynamic variation of archaeal community in AnMBRs was not as obvious as that of bacterial community based on the PCR-DGGE results. Under the circumstance of stable operation, Cloacimonetes, Chloroflexi, Bacteroidetes, Proteobacteria, Firmicutes, and Ignavibacteriae were observed as the predominant phyla in digested sludge based on the Illumina results. In addition to that, the cake layer possessed similar predominant phyla with the digested sludge but owned a higher diversity. Furthermore, overlapping bacterial communities were discovered between the excess sludge and digested sludge. However, the abundance of aerobic bacteria was substantially reduced, while the abundance of anaerobic microorganisms like phylum Cloacimonetes and Smithella was enriched in digested sludge over time. Additional PAC dosing, on the one hand, affected the bioavailable substrate, thus further changing the microbial community structure; on the other hand, aluminum itself also affected specific microbial communities. Besides, PAC dosing indirectly influenced the bacterial diversity in AnMBR as well.

Distance-Decay Relationship for Biological Wastewater Treatment Plants.

UNLABELLED: Patterns in the spatial distribution of organisms provide important information about mechanisms underlying biodiversity and the complexity of ecosystems. One of the most well-documented spatial patterns is the distance-decay relationship, which is a universal biogeographic pattern observed repeatedly for plant and animal communities, particularly for microorganisms in natural ecosystems such as soil, ocean, and salt marsh sediment. However, it is uncertain whether the microorganisms exhibit a distance-decay pattern in engineered ecosystems. Therefore, we measured the distance-decay relationship across various microbial functional and phylogenetic groups in 26 biological wastewater treatment plants (WWTPs) in China using a functional gene array (GeoChip 4.2). We found that microbial communities of activated sludge in WWTPs exhibited a significant but very weak distance-decay relationship. The taxon-area z values for different functional and phylogenetic groups were <0.0065, which is about 1 to 2 orders of magnitude lower than those observed in microbial communities elsewhere. Variation-partitioning analysis (VPA) showed that the relationships were driven by both environmental heterogeneity and geographic distance. Collectively, these results provided new insights into the spatial scaling of microbial communities in engineering ecosystems and highlighted the importance of environmental heterogeneity and geographic distance in shaping biogeographic patterns. IMPORTANCE: Determining the distance-decay relationship of microbial biodiversity is important but challenging in microbial ecology. All studies to date are based on natural environments; thus, it remains unclear whether there is such a relationship in an engineered ecosystem. The present study shows that there is a very weak distance-decay relationship in an engineered ecosystem (WWTPs) at the regional-to-continental scale. This study makes fundamental contributions to a mechanistic, predictive understanding of microbial biogeography.

Ammonia-oxidizing archaea versus bacteria in two soil aquifer treatment systems.

So far, the contribution of ammonia-oxidizing archaea (AOA) to ammonia oxidation in wastewater treatment processes has not been well understood. In this study, two soil aquifer treatment (SATs) systems were built up to treat synthetic domestic wastewater (column 1) and secondary effluent (column 4), accomplishing an average of 95% ammonia removal during over 550 days of operation. Except at day 322, archaeal amoA genes always outnumbered bacterial amoA genes in both SATs as determined by using quantitative polymerase chain reaction (q-PCR). The ratios of archaeal amoA to 16S rRNA gene averaged at 0.70 ± 0.56 and 0.82 ± 0.62 in column 1 and column 4, respectively, indicating that all the archaea could be AOA carrying amoA gene in the SATs. The results of MiSeq-pyrosequencing targeting on archaeal and bacterial 16S rRNA genes with the primer pair of modified 515R/806R indicated that Nitrososphaera cluster affiliated with thaumarchaeal group I.1b was the dominant AOA species, while Nitrosospira cluster was the dominant ammonia-oxidizing bacteria (AOB). The statistical analysis showed significant relationship between AOA abundance (compared to AOB abundance) and inorganic and total nitrogen concentrations. Based on the mathematical model calculation for microbial growth, AOA had much greater capacity of ammonia oxidation as compared to the specific influent ammonia loading for AOA in the SATs, implying that a small fraction of the total AOA would actively work to oxidize ammonia chemoautotrophically whereas most of AOA would exhibit some level of functional redundancy. These results all pointed that AOA involved in microbial ammonia oxidation in the SATs.

[Cultivation and characterization of an ammonia oxidizing archaeon enriched from wastewater treatment plant].

OBJECTIVES: To enrich ammonia-oxidizing archaeon (AOA) from wastewater treatment plants, identify its phylogenetic status, morphology and determine its growth and ammonia oxidation rates. METHODS: AOA was enriched in autotrophic medium containing antibiotics by using consecutive passage. The purity, uniformity and phylogenetic status of archaeal enrichment were determined with different molecular tools. The morphology was determined with scanning electron microscopy. The AOA growth andammonia oxidation rates were calculated from the corresponding experimental results. RESULTS: An AOA enrichment HJ-2b with purity of 93% was obtained. The similarity of its 16S rRNA gene with Nitrososphaera sp. JG1 was 100%, suggesting it belonged to Nitrososphaera spp., although the similarity of amoA gene between these two species was only 72%. HJ-2b cell was rod-shaped, with the maximum specific growth rate of 0. 43 d -1, the specific ammonia-oxidation rate of 3. 9 fmol/(cell . d). CONCLUSIONS: HJ-2b has great significance in studying the occurrence and contribution of AOA in wastewater treatment.

Overall functional gene diversity of microbial communities in three full-scale activated sludge bioreactors.

Understanding microbial community composition is thought to be crucial for improving process functioning and stabilities of full-scale activated sludge reactors in wastewater treatment plants (WWTPs). However, functional gene compositions of microbial communities within them have not been clearly elucidated. To gain a complete picture of microbial community, in this study, GeoChip 4.2 was used to profile the overall functional genes of three full-scale activated sludge bioreactors, the 16S rRNA gene diversities of which had been unveiled by 454-pyrosequencing in our previous investigation. Triplicate activated sludge samples from each system were analyzed, with the detection of 38,507 to 40,654 functional genes. A high similarity of 77.3-81.2 % shared functional genes was noted among the nine samples, verified by the high 16S rRNA gene similarity with shared operational taxonomic units (OTUs) constituting 66.4-70.0 % of the detected sequences in each system. Correlation analyses showed that the abundances of a wide array of functional genes were associated with system performances. For example, the abundances of carbon degradation genes were strongly correlated to chemical oxygen demand (COD) removal efficiencies (r = 0.8697, P < 0.01). Lastly, we found that sludge retention time (SRT), influent total nitrogen concentrations (TN inf), and dissolved oxygen (DO) concentrations were key environmental factors shaping the overall functional genes. Together, the results revealed vast functional gene diversity and some links between the functional gene compositions and microbe-mediated processes.

Continuous chain elongation process for carbon resource recovery from excess sludge: Enhanced n-caprylate production and specific microbial functionalities.

Thermal hydrolyzed sludge (THS) exhibits considerable promise in generating medium-chain fatty acids (MCFAs) through chain elongation (CE) technology. This study developed a novel continuous CE process using THS as the substrate, achieving an optimal ethanol loading rate (5.8 g COD/L/d) and stable MCFA production at 10.9 g COD/L, with a rate of 3.6 g COD/L/d. The MCFAs primarily comprised n-caproate and n-caprylate, representing 41.5 % and 54.3 % of the total MCFAs, respectively. Utilization efficiencies for ethanol and acetate were nearly complete at 100 % and 92.8 %, respectively. Key microbial taxa identified under these optimal conditions included Alcaligenes, SRB2, Sporanaerobacter, and Kurthia, which were instrumental in critical pathways such as the generation of acetyl-CoA, the initial carboxylation of acetyl-CoA, the fatty acid biosynthesis cycle, and energy metabolism. This research provides a theoretical and technical blueprint for converting waste sludge into valuable MCFAs, promoting sustainable waste-to-resource strategies.

Long-term occurrence, resistance risk and chaotic characteristics of antibiotic resistance genes in sludge anaerobic digestion system.

The long-term occurrence, dynamics and risk of antibiotic resistance genes (ARGs) in anaerobic digestion (AD) of excess sludge (ES) are not fully understood. Therefore, 13-month metagenomic monitoring was carried out in a full-scale AD plant. The highest ARG abundance and risk scores were observed in spring. AD achieved a 35 % removal rate for the total ARG abundance, but the risk score of AD sludge was not always lower than ES samples, because of the higher proportion of Rank I ARGs in AD sludge. ARGs showed less obvious patterns under linear models compared with microbial community, implying their chaotic dynamics, which was further confirmed by nonlinearity tests. Empirical dynamic modeling performed better than the autoregressive integrated moving average model for ARG dynamics, especially for those with simple and nonlinear dynamics. This study highlighted spring for its higher ARG abundance and risk, and recommended nonlinear models for revealing the dynamics of ARGs.

Mesoporous silicas synthesis and application for lignin peroxidase immobilization by covalent binding method.

Immobilization of enzymes on mesoporous silicas (MS) allows for good reusability. MS with two-dimensional hexagonal pores in diameter up to 14.13 nm were synthesized using Pluronic P123 as template and 1,3,5-triisopropylbenzene as a swelling agent in acetate buffer. The surface of MS was modified by the silanization reagents 3-aminopropyltriethoxysilane. Lignin peroxidase (LiP) was successfully immobilized on the modified MS through covalent binding method by four agents: glutaraldehyde, 1,4-phenylene diisothiocyanate, cyanotic chloride and water-soluble carbodiimide. Results showed that cyanotic chloride provided the best performance for LIP immobilization. The loaded protein concentration was 12.15 mg/g and the immobilized LiP activity was 812.9 U/L. Immobilized LiP had better pH stability. Acid Orange II was used to examine the reusability of immobilized LiP, showing more than 50% of the dye was decolorized at the fifth cycle.

Chemodiversity transformation of organic matters in a full scale MBR-NF wastewater reclamation plant.

Membrane bioreactor (MBR) and nanofiltration (NF) process has been attractive in wastewater reclamation, and was set as the target process in this study. Dissolved organic matter (DOM) and trace organic contaminants (TrOCs), closely associated with water safety, are noteworthy pollutants. Though the general DOM characteristics and TrOCs removal in MBR-NF reclamation process have been reported in lab-/pilot-scale experiment, the molecular characteristics of DOM revealed by high resolution mass spectrometry, and the correlation between DOM and TrOCs have been rarely studied in full-scale MBR-NF wastewater reclamation plant. In this work, biological and NF processes contributed significantly to the removal of DOM and TrOCs, while MBR filtration contributed slightly. Spectroscopic analyses revealed that DOM with higher aromaticity and lower molecular weight were more recalcitrant along the treatment. Aromatic protein-like substances were preferentially removed comparing to humic-like substances. Fourier transform ion cyclotron resonance mass spectrometry was applied to investigate DOM transformation at molecular level. DOM molecules with higher H/C and lower O/C, especially the aliphatics and peptides, were readily biodegraded into higher­oxygenate, highly unsaturated, and aromatic compounds. The generated species mainly included condensed aromatics, polyphenols, and highly unsaturated compounds. Filtration in MBR tended to reject higher oxygenated molecules. NF effectively removed most of the DOM molecules, especially higher oxygenated molecules with low H, N and S. The residual TrOCs in the NF effluent, including sulfamethoxazole, ofloxacin, and bisphenol A, still displayed above medium environmental risk. Significant correlations were found among organic compounds, spectral indices, and peptides molecules. Positive correlation between most of the TrOCs and several DOM parameters implied that they were synchronously removed in biological and membrane filtration processes. SUVA and FI might be potential indexes in monitoring the performance of MBR-NF process in both DOM and TrOC removal. These findings would expand the understanding of DOM and TrOCs behavior in wastewater reclamation process and simplify an in-depth system monitoring.

A hyperthermophilic anaerobic fermentation platform for highly efficient short chain fatty acids production from thermal hydrolyzed sludge.

In this study, a carboxylate platform of hyperthermophilic (70 â„ƒ) anaerobic fermentation (HAF) for short chain fatty acids (SCFAs) production from thermal hydrolyzed sludge (THS) was established. The long-term performance for SCFAs production and the microbial communities of this HAF under different SRTs were systematically investigated. Under the optimum SRT of 3 d, the HAF had the highest acetate production rate of 1.12 g COD/L/d which accounted for 60% in SCFAs. It also rendered a good performance in SCFAs production, with concentration, production rate and yield of 6.61 g COD/L, 1.86 g COD/L/d and 324 g COD/kg VSSin, respectively. Nearly no biogas produced from this system, which reduced the loss of carbon sources from the system. This was due to the inhibition of methanogenesis by the hyperthermophilic condition and the high content of total ammonia nitrogen (TAN) and free ammonia nitrogen (FAN). Tepidimicrobium, Bhargavaea and XBB1006 were the dominant genus-level biomarkers under the optimum SRT, which facilitated the decomposition of monosaccharides, amino acids, terpenoids and polyketides into SCFAs. This work provides an applicable anaerobic carboxylate platform for highly efficient SCFAs production from excess sludge.

Insight into the key role of oxygen dopants over ball-milled boron nitride for efficient degradation of PFOS alternative 6:2 fluorotelomer sulfonic acid.

6:2 Fluorotelomer sulfonic acid (6:2 FTS) has been identified as an alternative to perfluorooctane sulfonic acid but has been proven to cause potential threats to humans and the environment. In this study, boron nitride (BN) photocatalysis was explored for 6:2 FTS degradation with 100% removal (kobs=1.8 h-1) and desulfurization rate of 100% as well as the defluorination rate of 57.3%. The superior performance of BN was primarily related to oxygen dopants defects (O-dopants). In addition, O-dopants contribution was confirmed by ball-milled BN (B-BN), which introduced more O-dopants and exhibited an increased 6:2 FTS degradation rate of 2.88 h-1. The decomposition of 6:2 FTS was attributed to holes (h+), hydroxyl radicals (•OH), and superoxide (•O2-) and proceeded via two pathways, the hydrogen abstraction from ethyl carbons by •OH and the C-S bond activation by h+ and •OH. To the best of our knowledge, this is the first study demonstrating that h+, •OH, and •O2- played significant roles in the heterogeneous photocatalytic degradation of 6:2 FTS.

Spectroscopic fingerprints profiling the polysaccharide/protein/humic architecture of stratified extracellular polymeric substances (EPS) in activated sludge.

Extracellular polymeric substances (EPS), with a stratified structure including tightly-bound EPS (TB-EPS), loosely-bound EPS (LB-EPS), and soluble EPS (S-EPS) surrounding the microbial cells, are known to vitally affect the physicochemical and biological functions of activated sludge in wastewater treatment. Polysaccharides (PS), proteins (PN), and humic acids (HA) are key components of EPS but their roles in constructing the multi-layer architecture are still unclear. This study explored the EPS characteristics in relation to the components using spectroscopic fingerprinting techniques. Ultraviolet-visible (UV-vis) spectra demonstrated stark difference between TB-EPS and other EPS. Fluorescence excitation-emission matrix (FEEM) and apparent quantum yield revealed further detailed differences. Fluorescence quotient analysis highlighted the dominance of TB-EPS, LB-EPS, and S-EPS in the excitation/emission wavelength (Ex/Em) region of Em = 350-400 nm, Em > 400 nm, and low-Stokes shift band (Em - Ex < 25 nm), respectively. Wavelength-wise prediction of the FEEM intensity was achieved through multiple linear regression against the chemical composition and variance partitioning analysis witnessed binary interactions of PS×HA and PS×PN in S-EPS, PN×HA and PS×PN in LB-EPS, and ternary interaction of PS×PN×HA in TB-EPS as well as the wavelength-specific fluorescence responses of these interactions. Further, X-ray photoelectron spectroscopy, infrared spectra, and circular dichroism spectra corroborated the differences in primary, secondary, and tertiary structures across the EPS layers. Ultrahigh-performance liquid chromatography-mass spectrometry detected molecular fragments confirming the multi-component hybridization among PS, PN, and HA. This study demonstrates a spectroscopic approach to sensitively fingerprint the fine structure of EPS, which has the potential for rapid monitoring of EPS and related sludge properties in wastewater treatment systems.