Energy-Saving Mechanism of Wastewater Treatment Process Adaptation on Natural Temperature Variation: The Case from Coking Wastewater.

The cyclical variations in environmental temperature generated by natural rhythms constantly impact the wastewater treatment process through the aeration system. Engineering data show that fluctuations in environmental temperature cause the reactor temperature to drop at night, resulting in increased dissolved oxygen concentration and improved effluent wastewater quality. However, the impact of natural temperature variation on wastewater treatment systems and the energy-saving potential has yet to be fully recognized. Here, we conducted a comprehensive study, using a full-scale oxic-hydrolytic and denitrification-oxic (OHO) coking wastewater treatment process as a case and developed a dynamic aeration model integrating thermodynamics and kinetics to elucidate the energy-saving mechanisms of wastewater treatment systems in response to diurnal temperature variations. Our case study results indicate that natural diurnal temperature variations can cut the energy consumption of 660,980 kWh annually (up to 30%) for the aeration unit in the OHO system. Wastewater treatment facilities located in regions with significant environmental temperature variation stand to benefit more from this energy-saving mechanism. Methods such as flow dynamic control, load shifting, and process unit editing can be fitted into the new or retrofitted wastewater treatment engineering.

Activated carbon enhanced traditional activated sludge process for chemical explosion accident wastewater treatment.

With the development of industries, explosion accidents occur frequently during production, transportation, usage and storage of hazard chemicals. It remained challenging to efficiently treat the resultant wastewater. As an enhancement of traditional process, the activated carbon-activated sludge (AC-AS) process has a promising potential in treating wastewater with high concentrations of toxic compounds, chemical oxygen demand (COD) and ammonia nitrogen (NH4+-N), etc. In this paper, activated carbon (AC), activated sludge (AS) and AC-AS were used to treat the wastewater produced from an explosion accident in the Xiangshui Chemical Industrial Park. The removal efficiency was assessed by the removal performances of COD, dissolved organic carbon (DOC), NH4+-N, aniline and nitrobenzene. Increased removal efficiency and shortened treatment time were achieved in the AC-AS system. To achieve the same COD, DOC and aniline removal (90%), the AC-AS system saved 30, 38 and 58 h compared with the AS system, respectively. The enhancement mechanism of AC on the AS was explored by metagenomic analysis and three-dimensional excitation-emission-matrix spectra (3DEEMs). More organics, especially aromatic substances were removed in the AC-AS system. These results showed that the addition of AC promoted the microbial activity in pollutant degradation. Bacteria, such as Pyrinomonas, Acidobacteria and Nitrospira and genes, such as hao, pmoA-amoA, pmoB-amoB and pmoC-amoC, were found in the AC-AS reactor, which might have played important roles in the degradation of pollutants. To sum up, AC might have enhanced the growth of aerobic bacteria which further improved the removal efficiency via the combined effects of adsorption and biodegradation. The successful treatment of Xiangshui accident wastewater using the AC-AS demonstrated the potential universal characteristics of the process for the treatment of wastewater with high concentration of organic matter and toxicity. This study is expected to provide reference and guidance for the treatment of similar accident wastewaters.

Microbial community composition and function prediction involved in the hydrolytic bioreactor of coking wastewater treatment process.

The hydrolytic acidification process has a strong ability to conduct denitrogenation and increase the biological oxygen demand/chemical oxygen demand ratio in O/H/O coking wastewater treatment system. More than 80% of the total nitrogen (TN) was removed in the hydrolytic bioreactor, and the hydrolytic acidification process contributed to the provision of carbon sources for the subsequent nitrification process. The structure and diversity of microbial communities were elaborated using high-throughput MiSeq of the 16S rRNA genes. The results revealed that the operational taxonomic units (OTUs) belonged to phyla Bacteroidetes, Betaproteobacteria, and Alphaproteobacteria were the dominant taxa involved in the denitrogenation and degradation of refractory contaminants in the hydrolytic bioreactor, with relative abundances of 22.94 ± 3.72, 29.77 ± 2.47, and 18.23 ± 0.26%, respectively. The results of a redundancy analysis showed that the OTUs belonged to the genera Thiobacillus, Rhodoplanes, and Hylemonella in the hydrolytic bioreactor strongly positively correlated with the chemical oxygen demand, TN, and the removal of phenolics, respectively. The results of a microbial co-occurrence network analysis showed that the OTUs belonged to the phylum Bacteroidetes and the genus Rhodoplanes had a significant impact on the efficiency of removal of contaminants that contained nitrogen in the hydrolytic bioreactor. The potential function profiling results indicate the complementarity of nitrogen metabolism, methane metabolism, and sulfur metabolism sub-pathways that were considered to play a significant role in the process of denitrification. These results provide new insights into the further optimization of the performance of the hydrolytic bioreactor in coking wastewater treatment.

Immobilization of Phosphatidylserine by Ethanol and Lysozyme on the Cell Surface for Evaluation of Apoptosis-Like Decay in Activated-Sludge Bacteria.

Accurate determination of microbial viability can be crucial in microbe-dominated biosystems. However, the identification of metabolic decay in bacterial cells can be elaborate and difficult. We sought to identify apoptosis-like bacterial processes by using annexin V-fluorescein isothiocyanate (FITC) (AVF), a probe typically used to stain phosphatidylserine (PS) on exposed cell membranes. The bacterial cell wall provides a barrier that is responsible for low efficiency of direct PS staining of decayed bacterial cells. This can be overcome by pretreatment of the bacteria with 70% ethanol, which fixates the bacteria and preserves the PS status, combined with lysozyme treatment to hydrolyze the cell wall. That treatment improved the efficiency of AVF staining considerably, as shown for pure strains of an Ochrobactrum sp. and a Micrococcus sp. Using this method, decayed bacterial cells (induced by starvation) were more strongly stained, indicating externalization of PS to a greater extent than seen for cells harvested at logarithmic growth. A multispecies microbial sludge was artificially decayed by heat treatment or alternating anoxic-oxic treatment, which also induced increased AVF staining, again presumably via decay-related PS externalization. The method developed proved to be efficient for identification of bacterial decay and has potential for the evaluation of multispecies bacterial samples from sources like soil matrix, bioaerosol, and activated sludge.IMPORTANCE Since the externalization of phosphatidylserine (PS) is considered a crucial characteristic of apoptosis, we sought to identify apoptosis-like decay in bacterial cells by PS staining using AVF. We show that this is possible, provided the bacteria are pretreated with ethanol plus lysozyme to remove a physical staining barrier and preserve the original, decay-related externalization of PS. Our work suggests that PS externalization occurs in starved bacteria and this can be quantified with AVF staining, providing a measure of bacterial decay. Since PS is the common component of the lipid bilayer in bacterial cell membranes, this approach also has potential for evaluation of cell decay of other bacterial species.

Metabolic Traits of Candidatus Accumulibacter clade IIF Strain SCELSE-1 Using Amino Acids As Carbon Sources for Enhanced Biological Phosphorus Removal.

Despite recent evidence from full-scale plants suggesting that Candidatus Accumulibacter may be capable of using amino acids, this metabolic trait has never been confirmed in a bioreactor experiment. Here we show that an enriched culture of Ca. Accumulibacter clade IIF strain SCELSE-1 could metabolize 11 of 20 α-amino acids, with aspartate, glutamate, asparagine, and glutamine resulting in the highest phosphorus removal. The anaerobic uptake of aspartate and glutamate was achieved through a glutamate/aspartate-proton symporter fully powered by the proton motive force (PMF). Under anaerobic conditions aspartate was deaminized and routed into core carbon metabolic pathways to form polyhydroxyalkanoates (PHA). The lack of genes encoding NADH dependent isocitrate dehydrogenase in the Ca. Accumulibacter genome resulted in a kinetic barrier for glutamate to be channelled to the TCA cycle. Glutamate was stored as glutamate polymer. When amino acids (aspartate or glutamate) and acetate were supplied together, Ca. Accumulibacter took up both carbon sources simultaneously, with the uptake rate of each carbon source largely preserved. Overall energy savings (up to 17%) were achieved under mixed carbon scenarios, due to the ability of Ca. Accumulibacter to rearrange its anaerobic carbon metabolism based on the reducing power, PMF and ATP balance.

High Dissolved Oxygen Selection against Nitrospira Sublineage I in Full-Scale Activated Sludge.

A single Nitrospira sublineage I OTU was found to perform nitrite oxidation in full-scale domestic wastewater treatment plants (WWTPs) in the tropics. This taxon had an apparent oxygen affinity constant lower than that of the full-scale domestic activated sludge cohabitating ammonium oxidizing bacteria (AOB) (0.09 ± 0.02 g O2 m-3 versus 0.3 ± 0.03 g O2 m-3). Thus, nitrite oxidizing bacteria (NOB) may in fact thrive under conditions of low oxygen supply. Low dissolved oxygen (DO) conditions selected for and high aeration inhibited the NOB in a long-term lab-scale reactor. The relative abundance of Nitrospira sublineage I gradually decreased with increasing DO until it was washed out. Nitritation was sustained even after the DO was lowered subsequently. The morphologies of AOB and NOB microcolonies responded to DO levels in accordance with their oxygen affinities. NOB formed densely packed spherical clusters with a low surface area-to-volume ratio compared to the Nitrosomonas-like AOB clusters, which maintained a porous and nonspherical morphology. In conclusion, the effect of oxygen on AOB/NOB population dynamics depends on which OTU predominates given that oxygen affinities are species-specific, and this should be elucidated when devising operating strategies to achieve mainstream partial nitritation.

Investigation of the fate of heavy metals based on process regulation-chemical reaction-phase distribution in an A-O1-H-O2 biological coking wastewater treatment system.

Regulation mechanism of typical substances including OH-, CN-, SCN-, S2-, NH3 on the distribution of heavy metals was investigated in coking wastewater treatment plant with our self-designed Anaerobic-Oxic-Hydrolytic-Oxic (A-O1-H-O2) system through engineering data exposure and computational density functional theory (DFT) verification. The results showed that coking sludge had superior enrichment ability for heavy metals, especially for the sludge from the A and H tanks. The enrichment ratio of the 8 heavy metals including Cd, Pb, Ni, Zn, Cu, Hg, Cr and As in coking waste sludge was found to be 6232 (comparing to these in the influent wastewater of A-O1-H-O2 system). The distribution of 8 heavy metals was closely related to their chemical (precipitation and/or complexation) and biochemical reaction potential with OH-, CN-, SCN-, S2-, NH3 in the A-O1-H-O2 system. The regulation mechanism of these precipitation and/or complexation agents on heavy metals was confirmed by DFT calculation. The stable energy of complexes formed between typical compounds and common heavy metal ions follow the order: OH: Cu2+>Pb2+>Zn2+>Cd2+>Hg2+>Ni2+; S2-: Pb2+>Cu2+>Zn2+>Cd2+>Hg2+>Ni2+; CN-: Zn2+>Cu2+>Cd2+>Hg2+>Pb2+>Ni2+; SCN-: Zn2+>Cd2+>Pb2+>Hg2+>Cu2+>Ni2+; NH3: Cu2+>Zn2+>Cd2+>Pb2+>Hg2+>Ni2+, providing reference for the judgement of which metal ions were preferentially combined with the typical compounds in coking wastewater. The results of this paper indicated that the enrichment of heavy metal ions in coking wastewater can be achieved by process design combined with the control of operating conditions (dissolved oxygen, hydraulic retention time, sludge retention time and pH), basing on the nature of heavy metal ions. Finally, the separation and differential management of heavy metals can be achieved.

Simultaneous decarburization, nitrification and denitrification (SDCND) in coking wastewater treatment using an integrated fluidized-bed reactor.

There are two problems in biological treatment of coking wastewater (CWW): incapability of pre-anaerobic treatment to eliminate the toxicity in wastewater, and the lack of carbon source for subsequent denitrification in pre-aerobic treatment. To achieve simultaneous decarburization, nitrification and denitrification (SDCND) in CWW treatment, biological carrier materials was used to build an integrated fluidized-bed reactor (Reactor B, RB). A conventional fluidized-bed reactor (Reactor A, RA) was used as a control reactor under the same condition. The results showed that RB was more advantageous since its removal efficiencies of COD and TN were 90% and 87%, respectively, which were significantly higher than these in RA (82% and 45%), at a hydraulic retention time (HRT) of 60 h. Microelectrode measurement indicated that oxygen transfer was limited inside the carrier where the formation of a dissolved oxygen (DO) concentration gradient was observed. Microbial community analysis showed that the aerobic and anoxic microenvironments in RB promoted the co-existence of a wider variety of bacteria, thus achieving SDCND. These results indicated the integrated fluidized-bed reactor exhibited promising feasibility for simultaneous carbon and nitrogen removal in CWW treatment under the same aeration driven conditions. The SDCND process realized by fluidized-bed reactor provided a reference for the treatment of toxic industrial wastewater with high carbon to nitrogen ratio.

Residual chemical oxygen demand (COD) fractionation in bio-treated coking wastewater integrating solution property characterization.

The refractory nature of residual COD in bio-treated coking wastewater (BTCW) creates barriers for its further treatment and reclamation. It is necessary to fractionate the residual COD in BTCW associated with characterization of solution properties. In this paper, a stepwise process composed of membrane filtration, coagulation, adsorption and ozonation was proposed to fractionate residual COD in the BTCW, in which the COD was stepwise reduced to near zero. In addition, the correlation between COD and water quality indexes as well as solution properties were discussed together with a safety assessment of the water quality. Results showed that the residual COD fractionation percentage contributed by suspended solids, colloids, dissolved organics and reductive inorganic substances in the BTCW was 43.7%, 22.1%, 26.2% and 4.9%, respectively. By stepwise fractionating of these substances, the residual COD was reduced from 168.8 to 5.2 mg L-1, and the UV254 value decreased from 1.90 to 0.15 cm-1. In addition, the particle size of the dominant substances contributing to the residual COD was smaller than 450 nm. Among these substances, the hydrophobic fraction accounted for 78.66% (in the term of TOC). Three-dimensional excitation-emission matrix (3D-EEM) analysis showed that hydrophobic neutral substances (HON) were the main fluorescence constituent in the BTCW, which was highly removable by adsorption. The residual COD after adsorption was mainly composed of reductive inorganic substances. Apart from pursuit of high COD removal rates, more emphasis should be given to the removal of toxic COD. Correlations were observed between the residual COD and water quality indicators as well as solution properties, providing a guideline for optimized removal of residual COD in the BTCW. In summary, these results gave a referential information about the nature of residual COD in the BTCW for the selection of advanced treatment technologies and the management of water quality safety.

An Effective Design of Electrically Conducting Thin-Film Composite (TFC) Membranes for Bio and Organic Fouling Control in Forward Osmosis (FO).

The organic foulants and bacteria in secondary wastewater treatment can seriously impair the membrane performance in a water treatment plant. The embedded electrode approach using an externally applied potential to repel organic foulants and inhibit bacterial adhesion can effectively reduce the frequency of membrane replacement. Electrode embedment in membranes is often carried out by dispensing a conductor (e.g., carbon nanotubes, or CNTs) in the membrane substrate, which gives rise to two problems: the leaching-out of the conductor and a percolation-limited membrane conductivity that results in an added energy cost. This study presents a facile method for the embedment of a continuous electrode in thin-film composite (TFC) forward osmosis (FO) membranes. Specifically, a conducting porous carbon paper is used as the understructure for the formation of a membrane substrate by the classical phase inversion process. The carbon paper and the membrane substrate polymer form an interpenetrating structure with good stability and low electrical resistance (only about 1Ω/□). The membrane-electrode assembly was deployed as the cathode of an electrochemical cell, and showed good resistance to organic and microbial fouling with the imposition of a 2.0 V DC voltage. The carbon paper-based FO TFC membranes also possess good mechanical stability for practical use.

Effects of CeO2 nanoparticles on system performance and bacterial community dynamics in a sequencing batch reactor.

The effects of CeO2 nanoparticles (NPs) on the system performance and the bacterial community dynamics in a sequencing batch reactor (SBR) were investigated, along with the fate and removal of CeO2 NPs within the SBR. Significant impact was observed on nitrification; NH4+-N removal efficiency decreased from almost 100% to around 70% after 6 days of continuous exposure to 1.0 mg/L of CeO2 NPs, followed by a gradual recovery until a stable value of around 90% after 20 days. Additionally, CeO2 NPs also led to a significant increase in the protein content in the soluble microbial products, showing the disruptive effects of CeO2 NPs on the extracellular polymeric substance matrix and related activated sludge structure. Denaturing gradient gel electrophoresis analysis showed remarkable changes in the bacterial community structure in the activated sludge after exposure to CeO2 NPs. CeO2 NPs were effectively removed in the SBR mainly via sorption onto the sludge. However, the removal efficiency decreased from 95 to 80% over 30 days. Mass balance evaluation showed that up to 50% of the NPs were accumulated within the activated sludge and were removed with the waste sludge.

Direct and complete phosphorus recovery from municipal wastewater using a hybrid microfiltration-forward osmosis membrane bioreactor process with seawater brine as draw solution.

We report a hybrid microfiltration-forward osmosis membrane bioreactor (MF-FOMBR) for direct phosphorus recovery from municipal wastewater in the course of its treatment. In the process, a forward osmosis (FO) membrane and a microfiltration (MF) membrane are operated in parallel in a bioreactor. FO membrane rejects the nutrients (e.g., PO4(3-), Ca(2+), Mg(2+), etc.) and results in their enrichment in the bioreactor. The nutrients are subsequently extracted via the MF membrane. Phosphorus is then recovered from the nutrients enriched MF permeate via precipitation without addition of an external source of calcium or magnesium. The use of seawater brine as a draw solution (DS) is another novel aspect of the system. The process achieved 90% removal of total organic carbon and 99% removal of NH4(+)-N. 97.9% of phosphate phosphorus (PO4(3-)-P) was rejected by the FO membrane and enriched within the bioreactor. >90% phosphorus recovery was achieved at pH 9.0. The precipitates were predominantly amorphous calcium phosphate with a phosphorus content of 11.1-13.3%. In principal, this process can recover almost all the phosphorus, apart from that assimilated by bacteria for growth. Global evaluation showed an overall phosphorus recovery of 71.7% over 98 days.

Simultaneous decarbonization and phosphorus removal by Tetrasphaera elongata with glucose as carbon source under aerobic conditions.

Previous researches have recognized the vital role of Tetrasphaera elongata in enhanced biological phosphorus removal systems, but the underlying mechanisms remain under-investigated. To address this issue, this study investigated the metabolic characteristics of Tetrasphaera elongata when utilizing glucose as the sole carbon source. Results showed under aerobic conditions, Tetrasphaera elongata exhibited a glucose uptake rate of 136.6 mg/(L·h) and a corresponding phosphorus removal rate of 8.6 mg P/(L·h). Upregulations of genes associated with the glycolytic pathway and oxidative phosphorylation were observed. Noteworthily, the genes encoding the two-component sensor histidine kinase and response regulator transcription factor exhibited a remarkable 28.3 and 27.4-fold increase compared with the group without glucose. Since these genes play a pivotal role in phosphate-specific transport systems, collectively, these findings shed light on a potential mechanism for simultaneous decarbonization and phosphorus removal by Tetrasphaera elongata under aerobic conditions, providing fresh insights into phosphorus removal from wastewaters.

Iron-enhanced microscale laboratory aerated filters in the treatment of artificial mariculture wastewater: A study on nitrogen removal performance and the impact on microbial community structure.

This study investigates the nitrogen removal efficacy and microbial community dynamics in seawater aquaculture effluent treatment using three different substrate combinations of microscale laboratory aerated filters (MFs) - MF1 (LECA), MF2 (LECA/Fe-C), and MF3 (LECA/Pyrite). The findings indicated that the COD removal exceeded 95% across all MFs, with higher removal efficiencies in MF2 and MF3. In terms of nitrogen removal performance, MF2 exhibited the highest average nitrogen removal of 93.17%, achieving a 12.35% and 3.56% increase compared to MF1 (80.82%) and MF3 (89.61%), respectively. High-throughput sequencing analysis revealed that the Fe-C substrate significantly enhanced the diversity of the microbial community. Notably, in MF2, the salinophilic denitrifying bacterium Halomonas was significantly enriched, accounting for 42.6% of the total microbial community, which was beneficial for nitrogen removal. Moreover, an in-depth analysis of nitrogen metabolic pathways and microbial enzymes indicated that MF2 and MF3 possessed a high abundance of nitrification and denitrification enzymes, related to the high removal rates of NH4+-N and NO3--N. Therefore, the combination of LECA with iron-based materials significantly enhances the nitrogen removal efficiency from mariculture wastewater.

Effects of intensive chlorine disinfection on nitrogen and phosphorus removal in WWTPs.

The increased use of disinfection since the pandemic has led to increased effective chlorine concentration in municipal wastewater. Whereas, the specific impacts of active chlorine on nitrogen and phosphorus removal, the mediating communities, and the related metabolic activities in wastewater treatment plants (WWTPs) lack systematic investigation. We systematically analyzed the influences of chlorine disinfection on nitrogen and phosphorus removal activities using activated sludge from five full-scale WWTPs. Results showed that at an active chlorine concentration of 1.0 mg/g-SS, the nitrogen and phosphorus removal systems were not significantly affected. Major effects were observed at 5.0 mg/g-SS, where the nitrogen and phosphorus removal efficiency decreased by 38.9 % and 44.1 %, respectively. At an active chlorine concentration of 10.0 mg/g-SS, the nitrification, denitrification, phosphorus release and uptake activities decreased by 15.1 %, 69.5-95.9 %, 49.6 % and 100 %, respectively. The proportion of dead cells increased by 6.1 folds. Reverse transcriptional quantitative polymerase chain reaction (RT-qPCR) analysis showed remarkable inhibitions on transcriptions of the nitrite oxidoreductase gene (nxrB), the nitrite reductase genes (nirS and nirK), and the nitrite reductase genes (narG). The nitrogen and phosphorus removal activities completely disappeared with an active chlorine concentration of 25.0 mg/g-SS. Results also showed distinct sensitivities of different functional bacteria in the activated sludge. Even different species within the same functional group differ in their susceptibility. This study provides a reference for the understanding of the threshold active chlorine concentration values which may potentially affect biological nitrogen and phosphorus removal in full-scale WWTPs, which are expected to be beneficial for decision-making in WWTPs to counteract the potential impacts of increased active chlorine concentrations in the influent wastewater.

Blind spots of universal primers and specific FISH probes for functional microbe and community characterization in EBPR systems.

Fluorescence in situ hybridization (FISH) and 16S rRNA gene amplicon sequencing are commonly used for microbial ecological analyses in biological enhanced phosphorus removal (EBPR) systems, the successful application of which was governed by the oligonucleotides used. We performed a systemic evaluation of commonly used probes/primers for known polyphosphate-accumulating organisms (PAOs) and glycogen-accumulating organisms (GAOs). Most FISH probes showed blind spots and covered nontarget bacterial groups. Ca. Competibacter probes showed promising coverage and specificity. Those for Ca. Accumulibacter are desirable in coverage but targeted out-group bacteria, including Ca. Competibacter, Thauera, Dechlorosoma, and some polyphosphate-accumulating Cyanobacteria. Defluviicoccus probes are good in specificity but poor in coverage. Probes targeting Tetrasphaera or Dechloromonas showed low coverage and specificity. Specifically, DEMEF455, Bet135, and Dech453 for Dechloromonas covered Ca. Accumulibacter. Special attentions are needed when using these probes to resolve the PAO/GAO phenotype of Dechloromonas. Most species-specific probes for Ca. Accumulibacter, Ca. Lutibacillus, Ca. Phosphoribacter, and Tetrasphaera are highly specific. Overall, 1.4% Ca. Accumulibacter, 9.6% Ca. Competibacter, 43.3% Defluviicoccus, and 54.0% Dechloromonas in the MiDAS database were not covered by existing FISH probes. Different 16S rRNA amplicon primer sets showed distinct coverage of known PAOs and GAOs. None of them covered all members. Overall, 520F-802R and 515F-926R showed the most balanced coverage. All primers showed extremely low coverage of Microlunatus (<36.0%), implying their probably overlooked roles in EBPR systems. A clear understanding of the strength and weaknesses of each probe and primer set is a premise for rational evaluation and interpretation of obtained community results.

Two new clades recovered at high temperatures provide novel phylogenetic and genomic insights into Candidatus Accumulibacter.

Candidatus Accumulibacter, a key genus of polyphosphate-accumulating organisms, plays key roles in lab- and full-scale enhanced biological phosphorus removal (EBPR) systems. A total of 10 high-quality Ca. Accumulibacter genomes were recovered from EBPR systems operated at high temperatures, providing significantly updated phylogenetic and genomic insights into the Ca. Accumulibacter lineage. Among these genomes, clade IIF members SCELSE-3, SCELSE-4, and SCELSE-6 represent the to-date known genomes encoding a complete denitrification pathway, suggesting that Ca. Accumulibacter alone could achieve complete denitrification. Clade IIC members SSA1, SCUT-1, SCELCE-2, and SCELSE-8 lack the entire set of denitrifying genes, representing to-date known non-denitrifying Ca. Accumulibacter. A pan-genomic analysis with other Ca. Accumulibacter members suggested that all Ca. Accumulibacter likely has the potential to use dicarboxylic amino acids. Ca. Accumulibacter aalborgensis AALB and Ca. Accumulibacter affinis BAT3C720 seemed to be the only two members capable of using glucose for EBPR. A heat shock protein Hsp20 encoding gene was found exclusively in genomes recovered at high temperatures, which was absent in clades IA, IC, IG, IIA, IIB, IID, IIG, and II-I members. High transcription of this gene in clade IIC members SCUT-2 and SCUT-3 suggested its role in surviving high temperatures for Ca. Accumulibacter. Ambiguous clade identity was observed for newly recovered genomes (SCELSE-9 and SCELSE-10). Five machine learning models were developed using orthogroups as input features. Prediction results suggested that they belong to a new clade (IIK). The phylogeny of Ca. Accumulibacter was re-evaluated based on the laterally derived polyphosphokinase 2 gene, showing improved resolution in differentiating different clades.

Symbiotic virus-bacteria interactions in biological treatment of coking wastewater manipulating bacterial physiological activities.

Biological treatment is commonly used in coking wastewater (CWW) treatment. Prokaryotic microbial communities in CWW treatment have been comprehensively studied. However, viruses, as the critical microorganisms affecting microbial processes and thus engineering parameters, still remain poorly understood in CWW treatment context. Employing viromics sequencing, the composition and function of the viral community in CWW treatment were discovered, revealing novel viral communities and key auxiliary metabolic functions. Caudovirales appeared to be the predominant viral order in the oxic-hydrolytic-oxic (OHO) CWW treatment combination, showing relative abundances of 62.47 %, 56.64 % and 92.20 % in bioreactors O1, H and O2, respectively. At the family level, Myoviridae, Podoviridae and Siphoviridae mainly prevailed in bioreactors O1 and H while Phycodnaviridae dominated in O2. A total of 56.23-92.24% of novel viral contigs defied family-level characterization in this distinct CWW habitat. The virus-host prediction results revealed most viruses infecting the specific functional taxa Pseudomonas, Acidovorax and Thauera in the entire OHO combination, demonstrating the viruses affecting bacterial physiology and pollutants removal from CWW. Viral auxiliary metabolic genes (AMGs) were screened, revealing their involvement in the metabolism of contaminants and toxicity tolerance. In the bioreactor O1, AMGs were enriched in detoxification and phosphorus ingestion, where glutathione S-transferase (GSTs) and beta-ketoadipyl CoA thiolase (fadA) participated in biodegradation of polycyclic aromatic hydrocarbons and phenols, respectively. In the bioreactors H and O2, the AMGs focused on cell division and epicyte formation of the hosts, where GDPmannose 4,6-dehydratase (gmd) related to lipopolysaccharides biosynthesis was considered to play an important role in the growth of nitrifiers. The diversities of viruses and AMGs decreased along the CWW treatment process, pointing to a reinforced virus-host adaptive strategy in stressful operation environments. In this study, the symbiotic virus-bacteria interaction patterns were proposed with a theoretical basis for promoting CWW biological treatment efficiency. The findings filled the gaps in the virus-bacteria interactions at the full-scale CWW treatment and provided great value for understanding the mechanism of biological toxicity and sludge activity in industrial wastewater treatment.

Genomics and metabolic characteristics of simultaneous heterotrophic nitrification aerobic denitrification and aerobic phosphorus removal by Acinetobacter indicus CZH-5.

This study provides for the first time a systematic understanding of Acinetobacter indicus CZH-5 performance, metabolic pathway and genomic characteristics for aerobic nitrogen (N) and phosphorus (P) removal. Acinetobacter indicus CZH-5 showed promising performance in heterotrophic nitrification aerobic denitrification and aerobic phosphorus removal. Under optimal conditions, the maximum ammonia-N, total nitrogen and orthophosphate-P removal efficiencies were 90.17%, 86.33%, and 99.89%, respectively. The wide tolerance range suggests the strong environmental adaptability of the bacteria. The complete genome of this strain was reconstructed. Whole genome annotation was used to re-construct the N and P metabolic pathways, and related intracellular substance metabolic pathways were proposed. The transcription levels of related functional genes and enzyme activities further confirmed these metabolic mechanisms. N removal was achieved via the nitrification-denitrification pathway. Furthermore, CZH-5 exhibited significant aerobic P uptake, with phosphate diesters as the main species of intracellular P.

Integrated genomics provides insights into the evolution of the polyphosphate accumulation trait of Ca. Accumulibacter.

Candidatus Accumulibacter, a prominent polyphosphate-accumulating organism (PAO) in wastewater treatment, plays a crucial role in enhanced biological phosphorus removal (EBPR). The genetic underpinnings of its polyphosphate accumulation capabilities, however, remain largely unknown. Here, we conducted a comprehensive genomic analysis of Ca. Accumulibacter-PAOs and their relatives within the Rhodocyclaceae family, identifying 124 core genes acquired via horizontal gene transfer (HGT) at its least common ancestor. Metatranscriptomic analysis of an enrichment culture of Ca. Accumulibacter revealed active transcription of 44 of these genes during an EBPR cycle, notably including the polyphosphate kinase 2 (PPK2) gene instead of the commonly recognized polyphosphate kinase 1 (PPK1) gene. Intriguingly, the phosphate regulon (Pho) genes showed minimal transcriptions, pointing to a distinctive fact of Pho dysregulation, where PhoU, the phosphate signaling complex protein, was not regulating the high-affinity phosphate transport (Pst) system, resulting in continuous phosphate uptake. To prevent phosphate toxicity, Ca. Accumulibacter utilized the laterally acquired PPK2 to condense phosphate into polyphosphate, resulting in the polyphosphate-accumulating feature. This study provides novel insights into the evolutionary emergence of the polyphosphate-accumulating trait in Ca. Accumulibacter, offering potential advancements in understanding the PAO phenotype in the EBPR process.