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Activation of enzymes by monovalent cations (M+) is a widespread phenomenon in biology. Despite this, there are few structure-based studies describing the underlying molecular details. Thiolases are a ubiquitous and highly conserved family of enzymes containing both K+-activated and K+-independent members. Guided by structures of naturally occurring K+-activated thiolases, we have used a structure-based approach to engineer K+-activation into a K+-independent thiolase. To our knowledge, this is the first demonstration of engineering K+-activation into an enzyme, showing the malleability of proteins to accommodate M+ ions as allosteric regulators. We show that a few protein structural features encode K+-activation in this class of enzyme. Specifically, two residues near the substrate-binding site are sufficient for K+-activation: A tyrosine residue is required to complete the K+ coordination sphere, and a glutamate residue provides a compensating charge for the bound K+ ion. Further to these, a distal residue is important for positioning a K+-coordinating water molecule that forms a direct hydrogen bond to the substrate. The stability of a cation–π interaction between a positively charged residue and the substrate is determined by the conformation of the loop surrounding the substrate-binding site. Our results suggest that this cation–π interaction effectively overrides K+-activation, and is, therefore, destabilised in K+-activated thiolases. Evolutionary conservation of these amino acids provides a promising signature sequence for predicting K+-activation in thiolases. Together, our structural, biochemical and bioinformatic work provide important mechanistic insights into how enzymes can be allosterically activated by M+ ions.

The activated sludge of a biochemical unit (WLK_OD) and an advanced denitrification unit (WLK_AD) were collected from a municipal wastewater treatment plant (WWTP), in which the TN concentration of effluent was less than 1.5 mg·L

This study presents a successful treatment of biological acidic Se(IV)- and Cd(II)-containing wastewater via the SBR with limited carbon source (100 mg/L COD). Scanning electron microscopy coupled with energy dispersive X-ray spectroscopy (SEM-EDS), high solution transmission electron microscopy (HRTEM) and X-ray photoelectron spectrometer (XPS) results verified the formation of elemental Se and CdSe nanoparticles in the sludge. The abundance of genera in the microbial community gradually changed over the treatment phases depending on the Se(IV) and Cd(II) exposure with different influent COD concentrations. The taxa of Proteiniclasticum, Clostridium_sensu_stricto_12, Longilinea and Mycobacterium were dominant. Redundancy analysis (RDA) indicates that COD concentrations had the greatest impact on Zoogloea and Pseudomonas by promoting an increased abundance and decreased abundance, respectively. Overall, the results extended our understanding of the mechanisms and microbial community responding for the Se(IV) and Cd(II) removal under limited carbon availability in acidic wastewater.

In this study, spent mushroom substrates (SMSs) were fermented anaerobically at room temperature to gain liquid SMSs (LSMSs) that were used to remove nitrogen from the piggery wastewater with a low C/N ratio in a sequencing batch reactor (SBR) and solid SMSs (SSMSs) that were utilized to adsorb Pb2+ from Pb2+-containing wastewater in a fixed-bed reactor (FBR). After LSMSs supplement, the removal efficiency of both total nitrogen (TN) and NH+4-N increased from around 50% to 60–80%. High-throughput sequencing results presented an obvious change in microbial diversity, and some functional microorganisms like Zoogloea and Hydrogenophaga predominated to promote nitrogen removal. Pb2+ did not emerge from the effluent until 240 min with the corresponding concentration being less than 3 mg/L when using 30-day SSMSs as adsorbents, and it was demonstrated to be appropriate to use the Thomas model to predict Pb2+ sorption on SSMSs. Although various functional groups played a role in binding ions, the carboxyl group was proved to contribute most to Pb2+ adsorption. These results certified that the anaerobically fermented SMSs are decidedly suitable for wastewater treatment. HIGHLIGHTS Anaerobically fermented spent mushroom substrates (AFSMS) could be recycled entirely.; AFSMSs were suitable for the treatment of piggery wastewater with a low C/N.; AFSMSs had a much higher capacity to adsorb Pb2+ than native SMSs.

The paper assesses the impact of the duration of oxic and anoxic conditions on the selection of microbial populations in laboratory-scale integrated fixed-film sequencing batch reactors (IF-SBRs). IF-SBRs were fed anaerobically with glucose and sodium acetate based synthetic wastewater. Three IF-SBRs were operated with different lengths of oxic and anoxic periods, and a control IF-SBR was operated with continuous aeration during the react phase of the SBR cycle. Compact flocs were formed in the control bioreactor, whereas granular sludge developed rapidly within ten days in the IF-SBR operated with an extended anoxic phase (3.25 hours within the six-hour cycle). Filamentous bulking was observed in the IF-SBRs that were operated with relatively moderate anoxic periods. Confocal laser scanning microscopy of the granular sludge revealed that β-polysaccharide and amyloid adhesins were predominant in the periphery and core of the granules, respectively. 16S rRNA gene sequencing (Illumina MiSeq) revealed rapid selection and increases in the relative abundance of Flavobacterium, Caulobacter, Dechloromonas, Zoogloea, and bacterial genera related to the family Comamonadaceae, that were predominant in the granular sludge. The functional capabilities of the bacteria that were predominant in the granular sludge include quorum sensing, formation of storage polymers, and denitrification. Optimizing the length of oxic and anoxic conditions in sequencing batch reactors, and in the configuration in this study which was a hybrid system consisting of flocs, biofilm and granular sludge, is an important consideration in the selection of bacterial populations important to the development of microbial structures, and granular sludge in particular.

To enhance the stability and pollutant removal performance of aerobic granular sludge (AGS), four groups of AGS reactors with mesh screens of different pore sizes (R1 is the control reactor; R2, R3, and R4 have pore sizes of 1.5, 2.5, and 3.5 mm, respectively) were established in this study. The results showed that the granular sludge in the reactor with a 2.5 mm mesh screen (R3) has the highest mixed liquor suspended solids (MLSS) value of 8.2 ± 0.2 g L−1 and the lowest sludge volume index (SVI30) value of 30 mL g−1. At the same time, the mature granular sludge in R3 had a compact structure and excellent total nitrogen (TN) removal performance (84.9 ± 1.2%), along with an optimal granule size distribution (span value of 96 ± 0.12). Analysis of the sludge microbial community showed that R3 has a higher enrichment of functional microorganisms, such as Zoogloea spp. (50.5%) and Thauera spp. (16.1%). This is beneficial for improving its intercellular adhesion and denitrification performance. The granule size distribution results indicated that R3 screened the growing granules with its appropriate mesh aperture in terms of optimizing hydraulic shear, thus inhibiting the growth of filamentous bacteria and keeping the granule size within the optimal range. This created a favorable niche for these fast-growing microorganisms, eventually forming granules with good stability and high denitrification efficiency.

Based on the prevalence of combined antibiotics and heavy metals contamination in the aquatic environment, this study utilized a microbial approach to achieve simultaneous removal of nitrate (NO

The discrepancy of c‐di‐GMP and EDR proved that TB‐EPS served as important component in the structure and growth of denitrification biofilm, denitrifiers could effectively receive electrons from both acetate and EPS, which was beneficial to maintain relatively high denitrification capability at low temperature. Pseudomonas played an important role to maintain stable structure by EPS production under shock organic loading rate and low temperature conditions, and the specialized communities suggested that good nitrate removal property could be enhanced not only by the interaction of unique OTUs but also through the high bioactivity of dominant species., Summary Microbial community may respond to different adverse conditions and result in the variation of extracellular polymeric substances (EPS) in denitrification biofilm; this study discovered the role of EPS in accordance with the analysis of cyclic diguanylate (c‐di‐GMP) and electron equilibrium (EE) under low organic loading rate, shock organic loading rate and low temperature conditions. Good nitrate removal performance could be achieved under shock organic loading rate and low temperature conditions; however, owing to the low organic loading rate, the carbon source was preferentially utilized for biomass growth. Tightly bound EPS (TB‐EPS) contents progressively increased and facilitated cell adhesion and biofilm formation. The stable TB protein (TB‐PN) content in TB‐EPS built a cross‐linked network to maintain internal biofilm structure and led to the rapid biosynthesis of polysaccharides, which could further enhance microbial adhesion and improve nitrate removal. C‐di‐GMP played an important role in biomass retention and biofilm formation, based on the correlation analysis of c‐di‐GMP and EPS. TB polysaccharide (TB‐PS) contents presented a significant positive correlation with c‐di‐GMP content, microbial adhesion and biofilm stabilization was further enhanced through c‐di‐GMP regulation. In addition, a remarkable negative correlation between electron deletion rate (EDR) and TB‐PN and TB‐PS was discovered, and TB‐PS was required to serve as energy source to enhance denitrification according to EE analysis. Surprisingly, dynamic microbial community was observed due to the drastic community succession under low temperature conditions, and the discrepancy between the dominant species for denitrification was found under shock organic loading rate and low temperature conditions. The notable increase in bacterial strains Simlicispira, Pseudomonas and Chryseobacterium was conducive to biofilm formation and denitrification under shock organic loading rate, while Dechloromonas and Zoogloea dramatically enriched for nitrate removal under low temperature conditions. The high abundance of Dechloromonas improved the secretion of EPS through the downstream signal transduction, and the c‐di‐GMP conserved in Pseudomonas concurrently facilitated to enhance exopolysaccharide production to shock organic loading rate and low temperature conditions.

This study investigates the stability of a pure oxygen aeration-activated sludge system for petrochemical wastewater treatment under high organic concentration and non-steady food-to-microorganism (F/M) ratio conditions. Sludge settling characteristics maintained relatively stable conditions with an F/M ratio variation from 0.15 ± 0.04 to 0.33 ± 0.07 kg COD/kg MLSS⋅d, while the excess F/M ratio (0.44 ± 0.16 kg COD/kg MLSS⋅d) resulted in deterioration of the organic removal and sludge-water separation performances. Loosely bound extracellular polymeric substances (EPS) showed more significant effect on sludge settleability than the tightly bound EPS. The genus

A motile, Gram-stain-negative, rod-shaped bacterium, designated G-4-1-14T, was obtained from forest soil sampled at Gwanggyo mountain, Gyeonggi-do, Republic of Korea. Cells were colourless, aerobic, grew optimally at 28–35 °C and hydrolysed DNA and casein. Phylogenetic analysis based on its 16S rRNA gene sequence revealed that strain G-4-1-14T formed a lineage within the genus Zoogloea . The closest members were Zoogloea resiniphila ATCC 70068T (98.6 % sequence similarity), Zoogloea caeni EMB43T (98.2 %), Zoogloea oryzae A-7T (97.7 %), Zoogloea ramigera IAM 12136T (96.9 %) and Zoogloea oleivorans BucT (96.2 %). The major respiratory quinone was ubiquinone-8 and the principal polar lipids were phosphatidylethanolamine, phosphatidyl-N-methylethanolamine, diphosphatidylglycerol and phosphatidylglycerol. The predominant cellular fatty acids were summed feature 3 (iso-C15 :0 2-OH/C16 : 1 ω7c) and C16 : 0. The DNA G+C content was 65.9 mol%. The average nucleotide identity and digital DNA–DNA hybridization relatedness values between strain G-4-1-14T and other type strains were ≤81.6 and ≤24.9 %, respectively, which are below the species demarcation thresholds. Based on the results of phenotypic, phylogenetic and genomic analyses, strain G-4-1-14T represents a novel species in the genus Zoogloea , for which the name Zoogloea dura sp. nov. is proposed. The type strain is G-4-1-14T (=KACC 21618T=NBRC 114358T). In addition, we propose emendation of the genus Zoogloea and the species Zoogloea oryzae and Zoogloea ramigera .

As part of the research program “2018 Rapid screening and identification of freshwater microorganisms using MALDI-TOF/MS library” freshwater samples were collected from a branch of the Nakdong River. Almost 300 antibiotic-resistant bacterial strains were isolated from freshwater samples and subsequently identified by 16S rRNA gene sequencing. Seventeen strains among the isolates shared high 16S rRNA gene sequence similarity (>99.0%) with known species that were not previously recorded in Korea, and each of the isolates also formed a robust phylogenetic clade with the closest species. These species were phylogenetically diverse, belonging to four phyla, seven classes, 10 orders, and 13 genera. At the genus and class level, the previously unrecorded species belonged to Rhodovarius, Xanthobacter, and Shinella of the class Alphaproteobacteria; Ottowia, Simplicispira, and Zoogloea of Betaproteobacteria; Pseudomonas, Acinetobacter, and Shewanella of Gammaproteobacteria; Arcobacter of Epsilonproteobacteria; Sphingobacterium of Sphingobacteriia; Trichococcus of Bacilli; and Leucobacter of Actinobacteria. The previously unrecorded species were further characterized by examining their gram-staining, colony and cell morphology, biochemical properties, and phylogenetic position.

The research on gaseous VOCs biofilters has often concentrated on process optimization. However, the microbial community change upon operating conditions is not well understood. In this study, three lab-scale biofilters treating gaseous toluene were operated for 66 days with different inocula under changes in inlet loads and nitrogen sources. Three biofilters were inoculated with activated sludge, river sediment or microbial consortia, respectively. The microbial community differed a lot initially but gradually deviated toward similar structures with the same dominant microorganisms, i.e. Proteobacteria, Actinobacteria (phylum level) and Rhodococcus,Pseudomonas(genus level). Among three biofilters, the two biofilters inoculated with activated sludge and river sediment showed higher microbial diversity, better VOCs removal performance and higher metabolic activity. Higher relative abundance of Alcanivorax (3% compared with lower than 0.03%), Pimelobacte (0.05% compared with lower than 0.01%)were detected under low inlet load, and Zoogloea(0.1%), Alkaliphilus(0.2%) were detected when the inlet load was increased. the abundance of Pseudomonasdecreased from 14% to 2% when ammonia was used as nitrogen source instead of nitrate, meanwhile the abundance of Bacillus and Gordoniaincreased from 0.01% to 0.05% and 0.8% to 5.8% respectively. Some special organisms were observed i.e. the intestinal microorganism.

The microbial ecosystems of the sludge were characterized in terms of the core community structure, functional pathways, and functional redundancy through Illumina MiSeq sequencing and PICRUSt analysis on the activated sludge (AS) samples from an extended activated aeration process. Based on the identified OTU distribution, we identified 125 core community genera, including 3 abundant core genera and 21 intermittent abundant core genera. Putative genera Nitrosomonas, Nitrotoga, Zoogloea, Novosphingobium, Thermomonas, Amaricoccus, Tetrasphaera, Candidatus Microthrix, and Haliscomenobacter, which are associated with functions of nitrifying, denitrifying, phosphorus accumulating, and bulking and foaming, were found to present as the core community organisms in the AS sampled from the conventional extended aeration AS processes. The high-abundant nitrogen metabolic pathways were associated with nitrate reduction to ammonium (DNRA and ANRA), denitrification, and nitrogen fixation, while the ammonia oxidation-related genes (amo) were rarely annotated in the AS samples. Strict functional redundancy was not found with the AS ecosystem as it showed a high correlation between the community composition similarity and function similarity. In addition, the classified dominant core genera community was found to be sufficient to characterize the functionality of AS, which could invigorate applications of 16S rDNA MiSeq sequencing and PICRUSt for the prediction of functions of AS ecosystems.

Inspired by the controlled release technique of microcapsules, a novel carbon source using starch/polyvinyl alcohol (PVA) as the active material and a polyamide layer (PAL) as the shell coating on the surface of the carbon source was prepared and used for denitrification. Compared to the pure starch/PVA, the PAL-coated carbon source showed a lower chemical oxygen demand (COD) concentration (20 mg L−1) in effluent with excellent removal rate of NO3−-N exceeding 95% when the mass ratio of starch to PVA was 3 : 1. The high content of starch in the composite carbon source contributed to a decrease of denitrification cost. The carbon balance result indicated that the carbon release of the PAL-coated carbon source was mainly caused by microbial action (72.9%). Besides, little difference of microbial community was observed between pure starch/PVA and PAL-starch/PVA supported systems and the vast majority of biofilm species belonged to the six major phyla Proteobacteria, Bacteroidetes, Firmicutes, Planctomycetes, Chloroflexi and Actinobacteria, and Thauera and Zoogloea were the major genera which were denitrifying bacteria. In all, these results demonstrated that the PAL played a vital role in controlling COD in effluent without degrading the denitrification efficiency, as well as changing the structure of the microbial community, which paves a new way to fabricate a novel solid carbon source with low cost and excellent denitrification performance.

Network theory is widely used to understand microbial interactions in activated sludge and numerous other artificial and natural environments. However, when using correlation-based methods, it is not possible to identify the directionality of interactions within microbiota. Based on the classic Granger test of sequencing-based time-series data, a new Microbial Causal Correlation Network (MCCN) was constructed with distributed ecological interaction on the directed, associated links. As a result of applying MCCN to a time series of activated sludge data, we found that the hub species OTU56, classified as belonging the genus Nitrospira, was responsible for completing nitrification in activated sludge, and mainly interacted with Proteobacteria and Bacteroidetes in the form of amensal and commensal relationships, respectively. Phylogenetic tree suggested a mutualistic relationship between Nitrospira and denitrifiers. Zoogloea displayed the highest ncf value within the classified OTUs of the MCCN, indicating that it could be a foundation for activated sludge through forming the characteristic cell aggregate matrices into which other organisms embed during floc formation. Overall, the introduction of causality analysis greatly expands the ability of a network to shed a light on understanding the interactions between members of a microbial community.

The remediation of multiple pollutants in water, for instance, nitrate, heavy metals, and antibiotics is urgent and necessary for the global water resources protection. Herein, a modified loofah bioreactor was designed for simultaneous denitrification, manganese (Mn) oxidation, and tetracycline (TC) removal. The maximum removal efficiencies of NO3–-N (91.97%), Mn(II) (71.25%), and TC (57.39%) were achieved at a hydraulic retention time (HRT) of 9 h, Mn(II) concentration of 20 mg L-1, and TC concentration of 1 mg L-1. SEM and XRD were carried out to characterize the bioprecipitation in the operation of bioreactor. TC addition affected the gaseous denitrification products, dissolved organic matter, as well as reduced the OTU in the bioreactor. The Zoogloea were regarded as the dominant species in the microbial community and played an essential role in the operation of bioreactor. Metagenomic analysis proved the great potential for denitrification, manganese oxidation, and antibiotic removal of loofah bioreactor.

Background Bacterial floc formation plays a central role in the activated sludge (AS) process. The formation of AS flocs has long been known to require exopolysaccharide biosynthesis. We had demonstrated that both expolysaccharides and PEP-CTERM (a short C-terminal domain includes a near-invariant motif Pro-Glu-Pro (PEP)) proteins were required for floc-forming in Zoogloea resiniphila MMB, a dominant AS bacterium. However, the PEP-CTERM proteins are not encoded in the genome of AS bacterium Shinella zoogloeoides ATCC 19623 (formerly known as Zoogloea ramigera I-16-M) and other sequenced AS bacteria strains. The mechanism underlying floc formation of Shinella and related AS bacteria remained largely unclear. Results In this study, we have sequenced and annotated the complete genome of S. zoogloeoides ATCC 19623 (aka I-16-M), previously isolated in USA and treated as the neotype for the AS floc-forming bacterium Zoogloea ramigera I-16-M, and another AS strain XJ20 isolated in China. Mariner transposon mutagenesis had been conducted to isolate floc-forming-deficient mutants in the strain ATCC 19623 as previously performed by using Tn5 transposon three decades ago. The transposon insertional sites of multiple mutants were mapped to the gene cluster for bacterial cellulose synthesis (bcs) and secretion, and the role played by these genes in floc-formation had been further confirmed by genetic complementation. Interestingly, the restriction map of this bcs locus-flanking region was highly similar to that of the previously identified DNA fragment required for floc-formation in 1980s. Cellulase treatment abolished the floc-forming phenotype of S. zoogloeoides ATCC 19623 but not that of Z. resiniphila MMB strain. The FTIR spectral analyses revealed that the samples extracted from S. zoogloeoides ATCC 19623 were cellulose polymer. Conclusion Our results indicated that we have largely reproduced and completed the unfinished pioneering work on AS floc-formation mechanism, demonstrating that the floc-formation and flocculating capability of Shinella were mediated by extracellular cellulose polymers.

With the increasing occurrences of nitrate and Cr(VI) pollution globally, microbially driven pollutant reduction and its interaction effects were of growing interest. Despite the increasing number of experimental reports on the simultaneous reduction of nitrate and Cr(VI), a broad picture of the keystone species and metabolic differences in this process remained elusive. This study explored the changing of microorganisms with the introduction of Cr(VI)/NO3− through analyzing 242 samples from the NCBI database. The correlation between microbial abundance and environmental factors showed that, the types of energy substances and pollutants species in the environment had an impact on the diversity of microorganisms and community structure. The genus of Zoogloea, Candidatus Accumulibacter, and Candidatus Kapabacteria sp. 59–99 had the ability of denitrification, while genus of Alcaligenes, Kerstersia, Petrimonas, and Leucobacter showed effectively Cr(VI) resistance and reducing ability. Azoarcus, Pseudomonas, and Thauera were recognized as important candidates in the simultaneous reduction of nitrate and Cr(VI). Metagenomic predictions of these microorganisms using PICRUSt2 further highlighted the enrichment of Cr(VI)and nitrate reduction-related genes (such as chrA and norC). Special attention should therefore be paid to these bacteria in subsequent studies to evaluate their performance and mechanisms involved in simultaneous denitrification and chromium removal. The microbial co-occurrence network analysis conducted on this basis emphasized a strong association between community collaboration and pollution removal. Collectively, either site surveys or laboratory experiments, subsequent studies should focus on these microbial populations and the interspecific collaborations as they strongly influence the occurrence of simultaneous nitrate and Cr(VI) reduction.

A Mn(II) oxidizing-denitrifying and tetracycline (TC) removal bacterium Zoogloea sp. MFQ7 was isolated in this study. Nitrogen removal was 83.49% by nitrogen balance experiment. The maximum removal efficiencies of nitrate, Mn(II), and TC by strain MFQ7 within 96 h was 100.00, 74.56, and 63.59% at C/N of 2.0, pH of 7.0, Mn(II) of 20 mg L−1, temperature of 30.0 °C, and TC of 0.2 mg L−1. SEM illustrated that biogenic manganese oxides (BMO) was petal-like, XRD and XPS analyses confirmed that MnO2 was the main component of BMO. Besides, the maximum adsorption capacity of BMO for TC was 52.21 mg g−1. FTIR detected the changes in TC adsorption by BMO. Pseudo-second-order model (R2 = 0.994) explained the adsorption kinetics of TC on BMO and Langmuir isotherm model (R2 = 0.983) suggested that it was homogeneous adsorption, thermodynamics data (ΔG

Zoogloea oleivorans, capable of using toluene as a sole source of carbon and energy, was earlier found to be an active degrader under microaerobic conditions in aquifer samples. To uncover the genetic background of the ability of microaerobic toluene degradation in Z. oleivorans, the whole-genome sequence of the type strain BucT was revealed. Metatranscriptomic sequence reads, originated from a previous SIP study on microaerobic toluene degradation, were mapped on the genome. The genome (5.68 Mb) had a mean G + C content of 62.5%, 5005 protein coding gene sequences and 80 RNA genes. Annotation predicted that 66 genes were involved in the metabolism of aromatic compounds. Genome analysis revealed the presence of a cluster with genes coding for a multicomponent phenol-hydroxylase system and a complete catechol meta-cleavage pathway. Another cluster flanked by mobile-element protein coding genes coded a partial catechol meta-cleavage pathway including a subfamily I.2.C-type extradiol dioxygenase. Analysis of metatranscriptomic data of a microaerobic toluene-degrading enrichment, containing Z . oleivorans as an active-toluene degrader revealed that a toluene dioxygenase-like enzyme was responsible for the ring-hydroxylation, while enzymes of the partial catechol meta-cleavage pathway coding cluster were responsible for further degradation of the aromatic ring under microaerobic conditions. This further advances our understanding of aromatic hydrocarbon degradation between fully oxic and strictly anoxic conditions.

NaOH could promote volatile fatty acids (VFAs) with the deterioration of sludge dewaterability and high concentrations of ammonia nitrogen (NH4+-N) and soluble phosphorus (SP) in sludge fermentation system. Whether calcium peroxide (CaO2), as a solid base, could improve the status quo is unknown. Therefore it is necessary to study the effect of CaO2 on primary sludge (PS) fermentation. Five experimental groups were conducted with different gradient doses of CaO2 and high-throughput sequencing was conducted to reveal the mechanisms. The results showed that CaO2 contributed to enhancing the yield of organic matter, but also reduced SP and improved the sludge dewaterability. The optimal CaO2 dosage was 0.10g/g VSS, and the yields of VFAs elevated from 872 to 2794 mg COD L−1, respectively. The concentration of SP in fermentation liquor declined from 23.74 to 3.23 mg L−1. The sludge dewaterability was improved with decreasing capillary suction time (CST) from 365 to 227 s. NH4+-N in fermentation was also controlled and the concentration declined from 581 to 276 mg L−1. For revealing the mechanism, high-throughput sequencing results demonstrated that CaO2 enhanced the bacterial richness and diversity, and Alkaliphilus, Acinetobacter, Macellibacteroides, Arobacter and Zoogloea significant increased with CaO2 adding. Meanwhile, metabolic pathway abundance results demonstrated that CaO2 promoted the microbial metabolism, and amino acid and lipid metabolisms led to VFAs and SCOD concentration increasing. As a whole CaO2 changed the microbial composition and metabolisms, and then effected soluble substrate generation.

The formation and characteristics of aerobic granular sludge (AGS) under different operational conditions in a sequencing batch reactor (SBR), designed to treat Mustard tuber wastewater (MTW, characterized as saline wastewater), had been investigated in this study. Morphology and structure during granulation were determined using a microscope with digital camera and scanning electron microscope (SEM). Granules formed in the reactors could be classified as zoogloea granules with a clear boundary outline and filamentous granules with mycelia bestrewing boundary. Zoogloea granules, cultivated in reactor R1 and R2, were with higher density than filamentous granules, cultivated in reactor R3, and consequently had a higher settling velocity. Results showed that divalent metal ions such as Ca2+ and Mg2+ with phosphate in inflow could transform into precipitates, serving as crystal nucleus and carriers for granulation. Moreover, appropriate organic loading, hydrodynamic

The feasibility of rapidly controlling activated sludge bulking and accelerating aerobic sludge granulation was evaluated by adding intact aerobic granular sludge (AGS) to the bulking activated sludge (BAS) reactor. Two ratios of AGS to BAS (0.2 in the first reactor (R1), and 0.4 in the second reactor (R2)) were tested. The results indicate that the addition of AGS immediately improved the settling ability of BAS (sludge volume index at 30 min (SVI30) in R1 and R2 decreased from 173.1 mL/g to 130.8 and 91.3 mL/g, respectively) and gradually increased the biomass concentration (mixed liquor suspended solids (MLSS) in R1 and R2 increased to 4722 and 5190 mg/L, respectively), thus resolving the sludge bulking problem. Meanwhile, adding AGS not only promoted the BAS growth in aggregates, but also facilitated the selection of well-settling aggregates at an early stage. Consequently, the granulation process was significantly accelerated. The granulation time in R1 and R2 was 14 and 10 days, respectively, indicating that the higher ratio of AGS to BAS can result in the faster granulation. Partial nitrification could be maintained during the BAS granulation process when the initial inoculation of nitritation sludge was large enough. Additionally, the microbial community changed during the BAS granulation process. The genera Thauera and Zoogloea belonging to family Rhodobacteraceae were speculated to play an important role in the BAS granulation.

The performances of anaerobic-anoxic-oxic processes with (A2O-B) and without (A2O-C) beneficial microorganisms were compared to provide valuable insights on how they are affected by changes in the microbial biomass and community composition. Although the A2O-B process showed lower concentrations of mixed liquor suspended solids and mixed liquor volatile suspended solids than the A2O-C process under identical operating conditions, the A2O-B process was more effective for the removal of organic materials and nutrients compared with the A2O-C process. Furthermore, the compressibility and settleability of the activated sludge were significantly better in the A2O-B process than in the A2O-C process due to the enhanced decomposition of extracellular polymeric substances. These results indicated that the inoculation of beneficial microorganisms may increase the proportions of microorganisms in relation to the removal of organic materials, nutrients (i.e., Zoogloea, Dechloromonas, Nitrospira, and Nitrosomonas) and the reduction of the excess sludge (i.e., Proteobacteria and Bacteroidetes).

This study investigated functional dynamics of microbial community in response to different selection pressures, with a focus on denitrification. Suspended-biomass experiments demonstrated limited aerobic and relatively higher anoxic nitrate and nitrite reduction capabilities; the highest NO2-N and NO3-N removal rates were 1.3 ± 0.1 and 0.74 ± 0.01 in aerobic and 1.4 ± 0.05 and 3.4 ± 0.1 mg/L.h in anoxic media, respectively. Key potential denitrifiers were identified as: (i) complete aerobic denitrifiers: Dokdonella, Flavobacterium, and Ca. Accumulibacter; (ii) complete anoxic denitrifiers: Acinetobacter, Pseudomonas, Arcobacter, and Comamonas; (iii) incomplete nitrite denitrifier: Diaphorobacter (aerobic/anoxic), (iv): incomplete nitrate denitrifiers: Thauera (aerobic/anoxic) and Zoogloea (strictly-aerobic). Granular biomass removed 72 mg/L NH4-N with no NOx− accumulation. Heterotrophic nitrification and aerobic denitrification were proposed as the principal nitrogen removal pathway in granular reactors, potentially performed by two key organisms Thuaera and Flavobacterium. Biodiversity analysis suggested that the selection pressure of nourishment condition was the decisive factor for microbial selection and nitrogen removal mechanism.

Fluidized-carriers were supplemented into the aerobic tank of a full-scale wastewater treatment plant (WWTP) using an anaerobic/anoxic/aerobic (A2/O) system to improve the nitrogen removal efficiency in effluents. The effects of carrier supplementation on denitrification ability and the bacterial community structures were investigated over 10 months. The results showed that the average effluent concentration of total nitrogen (TN) was maintained at 9.46 ± 1.14 mg/L, which was lower than 15.17 ± 2.00 mg/L in the effluent without carrier supplementation, indicating that adding fluidized-carriers into the aerobic tank contributed to nitrogen removal efficiency. A thick biofilm was formed after 4 months, which provided a good anoxic-aerobic microenvironment to the microbes. Illumina sequencing analysis showed a higher bacterial diversity in the biofilm. The relative abundance of nitrifying bacteria, denitrifying bacteria, and aerobic denitrifying bacteria in the biofilms was 13.68–39%, 11.56–12.17%, and 9.76–12.50%, respectively, which was beneficial for nitrogen removal in the system. The most prevalent genera were Nitrospira, Bacillus, Thauera, Hyphomicrobium, Acinetobacter, Zoogloea, Pseudomonas, and Paracoccus, which can metabolize nitrogenous or aromatic compounds and were the major functional bacterial genera, suggesting that these organisms play key roles in biodegradation processes in the carrier-added A2/O wastewater treatment system.

A kind of gel beads loaded with iron–carbon powder (Fe–C), Zoogloea sp. L2, and riboflavin (VB2) were prepared through cross–linking of sodium alginate (SA) to establish an immobilized bioreactor. The optimal ratio of SA beads was adjusted by orthogonal experiment. The change of oxidation–reduction potential (ORP) and the concentration of Fe2+ and Fe3+ showed that the addition of VB2 as a redox mediator can promote denitrification. Under the optimal conditions (carbon to nitrogen (C/N) ratio = 2.0, pH = 7.0, and hydraulic retention time (HRT) = 8 h), the nitrate removal efficiency (NRE) of bioreactor reached 98.48% (1.99 mg L–1h−1). Furthermore, Fourier transform infrared spectrometer (FTIR), Fluorescence excitation–emission matrix (EEM), X–ray diffraction (XRD), and gas chromatography (GC) analysis revealed that the immobilization and denitrification of the immobilized bioreactor were excellent. High throughput sequencing also showed that Zoogloea played a vital role in nitrate removal.

Anaerobic wastewater treatment is a promising technology for refractory pollutant treatment. The nano zero-valent iron (nZVI) assisted anaerobic system could enhance contaminant removal. In this work, we added nZVI into an anaerobic system to investigate the effects on system performances and metabolic mechanism for chloramphenicol (CAP) wastewater treatment. As nZVI concentrations increased from 0 to 1 g/L, the CAP removal efficiency was appreciably improved from 46.5% to 99.2%, while the CH4 production enhanced more than 20 times. The enhanced CAP removal resulted from the enrichments of dechlorination-related bacteria (Hyphomicrobium) and other functional bacteria (e.g., Zoogloea, Syntrophorhabdus) associated with refractory contaminants degradation. The improved CH4 production was ascribed to the increases in fermentative-related bacteria (Smithella and Acetobacteroides), homoacetogen (Treponema), and methanogens. The increased abundances of anaerobic functional genes further verified the mechanism of CH4 production. Furthermore, the abundances of potential hosts of antibiotic resistance genes (ARGs) were reduced under high nZVI concentration (1 g/L), contributing to ARGs attenuation. This study provides a comprehensive analysis of the mechanism in metabolic performance enhancement and ARGs attenuation during nZVI-assisted anaerobic CAP wastewater treatment.

In recent years, the externalization of electrons as part of respiratory metabolic processes has been discovered in many different bacteria and some archaea. Microbial extracellular electron transfer (EET) plays an important role in many anoxic natural or engineered ecosystems. In this study, an anaerobic methane-converting microbial community was investigated with regard to its potential to perform EET. At this point, it is not well-known if or how EET confers a competitive advantage to certain species in methane-converting communities. EET was investigated in a two-chamber electrochemical system, sparged with methane and with an applied potential of +400 mV versus standard hydrogen electrode. A biofilm developed on the working electrode and stable low-density current was produced, confirming that EET indeed did occur. The appearance and presence of redox centers at −140 to −160 mV and at −230 mV in the biofilm was confirmed by cyclic voltammetry scans. Metagenomic analysis and fluorescence in situ hybridization of the biofilm showed that the anaerobic methanotroph ‘Candidatus Methanoperedens BLZ2’ was a significant member of the biofilm community, but its relative abundance did not increase compared to the inoculum. On the contrary, the relative abundance of other members of the microbial community significantly increased (up to 720-fold, 7.2% of mapped reads), placing these microorganisms among the dominant species in the bioanode community. This group included Zoogloea sp., Dechloromonas sp., two members of the Bacteroidetes phylum, and the spirochete Leptonema sp. Genes encoding proteins putatively involved in EET were identified in Zoogloea sp., Dechloromonas sp. and one member of the Bacteroidetes phylum. We suggest that instead of methane, alternative carbon sources such as acetate were the substrate for EET. Hence, EET in a methane-driven chemolithoautotrophic microbial community seems a complex process in which interactions within the microbial community are driving extracellular electron transfer to the electrode.

The evaluation of effluent wastewater quality mainly relies on the assessment of conventional bacterial indicators, such as fecal coliforms and enterococci; however, little is known about opportunistic pathogens, which can resist chlorination and may be transmitted in aquatic environments. In contrast to conventional microbiological methods, high-throughput molecular techniques can provide an accurate evaluation of effluent quality, although a limited number of studies have been performed in this direction. In this work, high-throughput amplicon sequencing was employed to assess the effectiveness of chlorination as a disinfection method for secondary effluents. Common inhabitants of the intestinal tract, such as Bacteroides, Arcobacter and Clostridium, and activated sludge denitrifiers capable of forming biofilms, such as Acidovorax, Pseudomonas and Thauera, were identified in the chlorinated effluent. Chloroflexi with dechlorination capability and the bacteria involved in enhanced biological phosphorus removal, i.e., Candidatus Accumulibacter and Candidatus Competibacter, were also found to resist chlorination. No detection of Escherichia indicates the lack of fecal coliform contamination. Mycobacterium spp. were absent in the chlorinated effluent, whereas toxin-producing cyanobacteria of the genera Anabaena and Microcystis were identified in low abundances. Chlorination significantly affected the filamentous bacteria Nocardioides and Gordonia, whereas Zoogloea proliferated in the disinfected effluent. Moreover, perchlorate/chlorate- and organochlorine-reducing bacteria resisted chlorination.

The pollution of multifarious pollutants such as heavy metal, organic compounds, and nitrate are a hot research topic at present. In this study, the functions of Zoogloea sp. MFQ7 and its biological precipitation formed during bacterial manganese oxidation on the removal of phenol and 17β-estradiol (E2) were investigated. Strain MFQ7, a manganese-oxidizing bacteria, can remove 98.34% of phenol under pH of 7.1, a temperature of 30 ℃ and Mn