Metagenomic Insight Reveals the Microbial Structure and Function of the Full-Scale Coking Wastewater Treatment System:Gene-Based Nitrogen Removal

Microbial communities play crucial roles in pollutant removal and system stability in biological systems for coking wastewater(CWW)treatment,but a comprehensive understanding of their structure and functions is still lacking.A five month survey of four sequential bioreactors,anoxic 1/oxic 1/anoxic 2/oxic 2(A1/O1/A2/O2),was carried out in a full-scale CWW treatment system in China to elucidate operational performance and microbial ecology.The results showed that A1/O1/A2/O2 had excellent and stable performance for nitrogen removal.Both total nitrogen(TN;(17.38±6.89)mgL1)and ammonium-nitrogen(NH4 t-N;(2.10±1.34)mg·L^(-1))in the final biological effluent satisfied the Chinese national standards for CWW.Integrated analysis of 16S ribosome RNA(rRNA)sequencing and metagenomic sequencing showed that the bacterial communities and metagenomic function profiles of A1 and O1 shared similar functional structures,while those of A2 significantly varied from those of other bioreactors(p<0.05).The results indicated that microbial activity was strongly connected with activated sludge function.Nitrosospira,Nitrosomonas,and SM1A02 were responsible for nitrification during the primary anoxic-oxic(AO)stage and Azoarcus and Thauera acted as important denitrifiers in A2.Nitrogen cycling-related enzymes and genes work in the A1/O1/A2/O2 system.Moreover,the hao genes catalyzing hydroxylamine dehydrogenase(EC 1.7.2.6)and the napA and napB genes catalyzing nitrate reductase(EC 1.9.6.1)played important roles in the nitrification and denitrification processes in the primary and secondary AO stages,respectively.The mixed liquor suspended solids(MLSS)/total solids(TS),TN removal rate(RR),total organic carbon(TOC)(RR),and NH_(4)^(+)t-N(RR)were the most important environmental factors for regulating the structure of core bacterial genera and nitrogen-cycling genes.Proteobacteria were the potential main participants in nitrogen metabolism in the A1/O1/A2/O2 system for CWW treatment.This study provides an original and comprehensive understanding of the microbial community and functions at the gene level,which is crucial for the efficient and stable operation of the full-scale biological process for CWW treatment.

Pretreatment of coking wastewater using anaerobic sequencing batch reactor (ASBR)

A laboratory-scale anaerobic sequencing batch reactor (ASBR) was used to pretreat coking wastewater. Inoculated anaerobic granular biomass was acclimated for 225 d to the coking wastewater, and then the biochemical methane potential (BMP)of the coking wastewater in the acclimated granular biomass was measured. At the same time, some fundamental technological factors, such as the filling time and the reacting time ratio (tf/tr), the mixing intensity and the intermittent mixing mode, that affect anaerobic pretreatment of coking wastewater with ASBR, were evaluated through orthogonal tests. The COD removal efficiency reached 38％～50％ in the stable operation period with the organic loading rate of 0.37～0.54 kg COD/(m3.d) at the optimum conditions of tf/tr, the mixing intensity and the intermittent mixing mode. In addition, the biodegradability of coking wastewater distinctly increased after the pretreatment using ASBR. At the end of the experiment, the microorganism forms on the granulated sludge in the ASBR were observed using SEM (scanning electron microscope) and fluoroscope. The results showed that the dominant microorganism on the granular sludge was Methanosaeta instead of Methanosarcina dominated on the inoculated sludge.

The Kinetics for Electrochemical Removal of Ammonia in Coking Wastewater

Electrochemical removal of ammonia is a new and effective method in coking wastewater.The reaction mechanism of ammonia removal was proved by stable polarization curve in this paper.First,the supposing of reaction steps of the electrode were proposed.And then reaction parameter of the electrode was measured by Tafel curve.Finally,the reaction mechanism was determined by quasi-equilibrium approach.The results showed that Cl2+H2O→HOCl+H++Cl was the rate-determining step,the calculated apparent transfer coefficient was uniform to the experimental value.

Advanced treatment of biologically pretreated coking wastewater by intensified zero-valent iron process(IZVI) combined with anaerobic filter and biological aerated filter(AF/BAF)

Experiments were conducted to investigate the behavior of the sequential system of intensified zero-valent iron process(IZVI) and anaerobic filter and biological aerated filter(AF/BAF) reactors for advanced treatment of biologically pretreated coking wastewater. Particular attention was paid to the performance of the integrated system for the removal of chemical oxygen demand(COD), ammonia nitrogen(NH3-N) and total nitrogen(TN). The average removal efficiencies of COD, NH3-N and TN were 76.28%, 96.76% and 59.97%, with the average effluent mass concentrations of 56, 0.53 and 18.83 mg/L, respectively, reaching the first grade of the national discharge standard. Moreover, the results of gas chromatography/mass spectrum(GC/MS) and gel permeation chromatography(GPC) analysis demonstrated that the refractory organic compounds with high relative molecular mass were partly removed in IZVI process by the function of oxidation-reduction, flocculation and adsorption which could also enhance the biodegradability of the system effluent. The removal efficiencies of NH3-N and TN were achieved mainly in the subsequent AF/BAF reactors by nitrification and denitrification. Overall, the results obtained show that the application of IZVI in combination with AF/BAF is a promising technology for advanced treatment of biologically pretreated coking wastewater.

Degradation analysis of A^2/O combined with AgNO3 + K2FeO4 on coking wastewater

In this work, a coking wastewater was selected and a biochemical Az/O treatment device for fractional degradation was designed and employed. After each stage of the treatment, the products were analyzed through gas chromatography-mass spectroscopy （GC-MS） to determine their composition. Finally, AgNO3 ＋ K2FeO4 was used as an advanced deep catalytic oxidation treatment. It was concluded from the analysis that cyclic organics could be degraded and the chemical oxygen demand （COD） was controlled within 50 mg. L-1, in line with the target value, Meanwhile, the spectra obtained from the GC-MS were in accordance with the conclusions reached based on the COD. The research results showed that all hard-degradable organics in coking wastewater could be eliminated through the A2/O bio-membrane treatment and the advanced treatment of making use of K2FeO4 as an oxidant and Ag＋ as a catalyst, the catalytic efficiency with Ag＋ as a catalyst of K2FeO4 was very high. Ag＋ could evidently improve the oxidation capacity of K2FeO4 to wastewater in its short stability time, and this is an important innovation.

Electrochemical treatment of COD in biologically pretreated coking wastewater using Ti/RuO2-IrO2 electrodes combined with modified coke

The electrochemical treatment of COD contained in biologically pretreated coking wastewater treated by a three-dimensional electrode system with modified coke as the particle electrode was investigated. And the electrochemical perromance of the coke modified with various active components was studied. The results show that the coke modified with Fe（NO3）2 has the lowest energy consumption and higher COD removal rate under the same condition, and the modified coke has better surface characteristics for the purpose of this study. In addition, the kinetic constant was also calculated. The study shows that the three-dimensional electrode system with Fe （NO3）z-modified coke can give a satisfactory solution in biologically pretreated coking wastewater.

Dynamics study on effect of temperature to Nitrous nitrification reaction of coking wastewater

Dynamic effects of NO2--N accumulation were discussed owing to temperature.In different temperature,a series of vmax and Ks were found considering the relation between the temperature and rate of ammonia nitrogen transforming into NO2--N.The kinetics models,which reflected the conditions of ammonia nitrogen transforming into NO2--N in the treatment process of the coking wastewater,were built up.The characteristic coefficient temperature was determined according to Arrhenius.

The influence of nitrified supernate’s recycle ratio on the removal of coking wastewater

The influence of the recycle ratio on the removal of coking wastewater has been researched using the anaerobicanoxic-aerobic （A/A/O） biofilm process. The research indicates that the concentrations of chemical oxygen demand （COD） and NO3 -N in the water are the lowest when the recycle ratio is 3：1 ,but the removal efficiency of total cyanide （TCN） is the highest when the recycle ratio is 1： 1. The removal efficiency of NH4^＋ -N is more than 99% at all three different recycle ratios. Compared with Grade A of the National Discharge Standard （GB 8978--1996）, the effluent NH4 -N is standard,but COD and TCN can not meet the requirements and further treatment processes are needed.

Temporal assembly patterns of microbial communities in three parallel bioreactors treating low-concentration coking wastewater with differing carbon source concentrations

Carbon source is an important factor of biological treatment systems, the effects of which on their temporal community assembly patterns are not sufficiently understood currently.In this study, the temporal dynamics and driving mechanisms of the communities in three parallel bioreactors for low-concentration coking wastewater(CWW) treatment with differing carbon source concentrations(S0 with no glucose addition, S1 with 200 mg/L glucose addition and S2 with 400 mg/L glucose addition) were comprehensively studied. Highthroughput sequencing and bioinformatics analyses including network analysis and Infer Community Assembly Mechanisms by Phylogenetic bin-based null model(iCAMP) were used. The communities of three systems showed turnover rates of 0.0029~0.0034 every 15days. Network analysis results showed that the S0 network showed higher positive correlation proportion(71.43%) and clustering coefficient(0.33), suggesting that carbon source shortage in S0 promoted interactions and cooperation of microbes. The neutral community model analysis showed that the immigration rate increased from 0.5247 in S0 to 0.6478in S2. The iCAMP analysis results showed that drift(45.89%) and homogeneous selection(31.68%) dominated in driving the assembly of all the investigated microbial communities.The contribution of homogeneous selection increased with the increase of carbon source concentrations, from 27.92% in S0 to 36.08% in S2. The OTUs participating in aerobic respiration and tricarboxylic acid(TCA) cycle were abundant among the bins mainly affected by deterministic processes, while those related to the metabolism of refractory organic pollutants in CWW such as alkanes, benzenes and phenols were abundant in the bins dominated by stochastic processes.

The significance of resource recycling for coking wastewater treatment:based on environmental and economic life cycle assessment

The sustainability of the coking industry is supported by reasonable production profit and environmental quality requirements.The traditional measures substantially increased the related costs for enterprises to reach standards.This paper aims to develop a comprehensive cost combined environmental impact assessment method that is necessary for the analysis of wastewater treatment systems.Typical three coking wastewater treatment processes in China were evaluated.Results showed that eutrophication dominantly contributed to the overall environmental effect.Improving effluent quality could significantly reduce the total environmental impact.In terms of an economic perspective,the price of raw materials was the main factor that affected the operating cost of comprehensive treatment.Based on subsystem analysis,the pretreatment stage accounted for the majority of environmental and cost burdens,respectively reaching 64%-78%and 64%-86%.Optimizing the pretreatment process by enhancing the efficiency of high concentration raw material recovery and substituting toxic raw materials for extractant could reduce the environmental impact and economic cost by 43.8%and 57%,respectively,which was an effective way to improve the potential performance of coking wastewater treatment plants(WWTPs).

Coking wastewater treatment for industrial reuse purpose: Combining biological processes with ultrafiltration, nanofiltration and reverse osmosis

A full-scale plant using anaerobic, anoxic and oxic processes （A1/A2/O）, along with a pilot-scale membrane bioreactor （MBR）, nanofiltration （NF） and reverse osmosis （RO） integrated system developed by Shanghai Baosteel Chemical Co. Ltd., was investigated to treat coking wastewater for industrial reuse over a period of one year. The removals reached 82.5% （COD）, 89.6% （BOD）, 99.8% （ammonium nitrogen）, 99.9% （phenol）, 44.6% （total cyanide （T-CN））, 99.7% （thiocyanide （SCN-）） and 8.9% （fluoride）, during the A1/A2/O biological treatment stage, and all parameters were further reduced by over 96.0%, except for fluoride （86.4%）, in the final discharge effluent from the currently operating plant. The pilot-scale MBR process reduced the turbidity to less than 0.65 NTU, and most of the toxic organic compounds were degraded or intercepted by the A1/A2/O followed MBR processes. In addition, parameters including COD, T-CN, total nitrogen, fluoride, chloride ion, hardness and conductivity were significantly reduced by the NF-RO system to a level suitable for industrial reuse, with a total water production ratio of 70.7%. However, the concentrates from the NF and RO units were highly polluted and should be disposed of properly or further treated before being discharged.

Removal of polycyclic aromatic hydrocarbons and phenols from coking wastewater by simultaneously synthesized organobentonite in a one-step process

The optimal condition for a one-step process removing organic compounds from coking wastewater by simultaneously synthesized organobentonite as a pretreatment was investigated.Results showed that sorption of organic compounds by organobentonite was positively correlated to the cation surfactant exchange on the bentonite and the octanol-water partition coefficient（Kow） of the solutes.With 0.75 g/L bentonite and 180 mg/L（60% of bentonite cation exchange capacity） cetyltrimethylammonium bromide,the removal efficiencies of the 16 polycyclic aromatic hydrocarbon（PAHs） specified by the US Environmental Protection Agency in coking wastewater except naphthalene were more than 90%,and that of benzo（a）pyrene was 99.5%.At the same time,the removal efficiencies of CODCr,NH3-N,volatile phenols,colour and turbidity were 28.6%,13.2%,8.9%,55% and 84.3%,respectively,and the ratio of BOD5/CODCr increased from 0.31 to 0.41.These results indicated that the one-step process had high removal efficiency for toxic and refractory hydrophobic organic compounds,and could improve the biodegradability of the coking wastewater.Therefore it could be a promising technology for the pretreatment of toxic and refractory organic wastewater.

Coking wastewater increases micronucleus frequency in mouse in vivo via oxidative stress

Coking wastewater has caused serious health risk in coal-producing areas of China, however its toxic effects have not been well understood. The genotoxicity induced by coking wastewater on mice in vivo and its possible oxidative mechanisms were investigated via observing the induction of micronuclei in polychromatic erythrocytes of mouse bone marrow, and subsequently determining the antioxidative enzyme activities （superoxide dismutase Cu, Zn-SOD, Se-dependent glutathione peroxidase, and catalase）, thiobarbituric acid reactive substance contents and protein carbonyl levels in brains and livers of mice. Results showed that the tested coking wastewater caused a significant increase of micronucleus frequencies in a concentration-dependent manner. Also, the sample increased lipid peroxidation and protein oxidation levels, which was accompanied by changes in antioxidative status. Interestingly, pre-treatment with an antioxidant （vitamin C） led to a statistical reduction in the micronucleus frequency caused by coking wastewater. This implies that coking wastewater induces evident genetic damage in mammalian cells, and exposure to polluted areas might pose a potential genotoxic risk to human beings; in the process, oxidative stress played a crucial role.

Revealing the anaerobic acclimation of microbial community in a membrane bioreactor for coking wastewater treatment by Illumina Miseq sequencing

The dynamic change of microbial community during sludge acclimation from aerobic to anaerobic in a MBR for coking wastewater treatment was revealed by Illumina Miseq sequencing in this study. The diversity of both Bacteria and Archaea showed an increase–decrease trajectory during acclimation, and exhibited the highest at the domestication interim. Ignavibacteria changed from a tiny minority（less than 1%） to the dominant bacterial group（54.0%） along with acclimation. The relative abundance of Betaproteobacteria kept relatively steady, as in this class some species increased coupled with some other species decreased during acclimation. The dominant Archaea shifted from Halobacteria in initial aerobic sludge to Methanobacteria in the acclimated anaerobic sludge. The dominant bacterial and archaeal groups in different acclimation stages were indigenous microorganisms in the initial sludge, though some of them were very rare. This study supported that the species in＂rare biosphere＂ might eventually become dominant in response to environmental change.

Biological removal of antiandrogenic activity in gray wastewater and coking wastewater by membrane reactor process

A recombinant human androgen receptor yeast assay was applied to investigate the occurrence of antiandrogens as well as the mechanism for their removal during gray wastewater and coking wastewater treatment. The membrane reactor（MBR） system for gray wastewater treatment could remove 88.0% of antiandrogenic activity exerted by weakly polar extracts and 97.3% of that by moderately strong polar extracts, but only 32.5%of that contributed by strong polar extracts. Biodegradation by microorganisms in the MBR contributed to 95.9% of the total removal. After the treatment, the concentration of antiandrogenic activity in the effluent was still 1.05 μg flutamide equivalence（FEQ）/L, 36.2%of which was due to strong polar extracts. In the anaerobic reactor, anoxic reactor, and membrane reactor system for coking wastewater treatment, the antiandrogenic activity of raw coking wastewater was 78.6 mg FEQ/L, and the effluent of the treatment system had only 0.34 mg FEQ/L. The antiandrogenic activity mainly existed in the medium strong polar and strong polar extracts. Biodegradation by microorganisms contributed to at least 89.2%of the total antiandrogenic activity removal in the system. Biodegradation was the main removal mechanism of antiandrogenic activity in both the wastewater treatment systems.

Pulsed corona discharge for improving treatability of coking wastewater

Coking wastewater（CW） contains toxic and macromolecular substances that inhibit biological treatment. The refractory compounds remaining in biologically treated coking wastewater（BTCW） provide chemical oxygen demand（COD） and color levels that make it unacceptable for reuse or disposal. Gas-phase pulsed corona discharge（PCD） utilizing mostly hydroxyl radicals and ozone as oxidants was applied to both raw coking wastewater（RCW） and BTCW wastewater as a supplemental treatment. The energy efficiency of COD,phenol, thiocyanate and cyanide degradation by PCD was the subject of the research. The cost-effective removal of intermediate oxidation products with addition of lime was also studied. The energy efficiency of oxidation was inversely proportional to the pulse repetition frequency： lower frequency allows more effective utilization of ozone at longer treatment times. Oxidative treatment of RCW showed the removal of phenol and thiocyanate at 800 pulses per second from 611 to 227 mg/L and from 348 to 86 mg/L, respectively, at 42 k Wh/m^3 delivered energy, with substantial improvement in the BOD5/COD ratio（from 0.14 to 0.43）.The COD and color of BTCW were removed by 30% and 93%, respectively, at 20 k Wh/m^3,showing energy efficiency for the PCD treatment exceeding that of conventional ozonation by a factor of 3–4. Application of lime appeared to be an effective supplement to the PCD treatment of RCW, degrading COD by about 28% at an energy input of 28 k Wh/m3 and the lime dose of 3.0 kg/m^3. The improvement of RCW treatability is attributed to the degradation of toxic substances and fragmentation of macromolecular compounds.

Removal of F- and organic matter from coking wastewater by coupling dosing FeCl3 and AlCl3

Coagulation and precipitation is a widely applied method to remove F-from wastewater.In this work,the effect of coagulation on the removal of F-and organic matter from coking wastewater was studied using Al Cl3and Fe Cl3as compound coagulants.The removal rates of F-and organic matter under different coagulant doses and p H conditions were investigated.The results show that the highest removal rates of F-by Al Cl3and Fe Cl3are 94.4%and 25.4%,respectively;when the dosage is 10 mmol/L,the TOC removal rates of Fe Cl3and Al Cl3reach 20.4%and 34.7%,respectively.Therefore,the removal rate of F-by Al Cl3is higher than that of Fe Cl3,but the removal rate of organic matter by Fe Cl3is relatively higher.The addition of Ca2+can promote the removal of F-,but the removal rate of organic matter decreases.In addition,by investigating the effects of different p H and Fe–Al ratio on the removal rate,the removal effect of adding Fe Cl3and Al Cl3at the same time was discussed.The results show that the most suitable working condition for the removal of organic matter and F-is that the p H is 6.5 and the molar ratio of Al/Fe is 8:2.Overall,the removal mechanism of F-and organic matter in coking wastewater by Fe Cl3and Al Cl3was explored in this study.The experimental results can provide reference for the advanced treatment of coking wastewater.

Preparation of Ti-based Yb-doped SnO_(2)-RuO_(2) electrode and electrochemical oxidation treatment of coking wastewater

In this study,the Ti/SnO_(2)-RuO_(2) electrodes with different Yb contents were prepared by sol-gel method and thermal decomposition method,and the surface morphology and crystal structure of the electrodes were characterized by scanning electron microscopy(SEM),atomic force microscopy(AFM) and X-ray diffraction(XRD),the electrochemical properties of the electrodes were tested by linear sweep voltammetry(LSV) and cyclic voltammetry(CV).The electrochemical oxidation device was constructed with Yb-doped Ti/SnO_(2)-RuO_(2) electrode as the anode and titanium plate as the cathode,and the electrochemical oxidation effect and product changes of the anode on co king wastewater were investigated.The results show that the surface of the electrode is flat with high crystallinity of SnO_(2) and RuO_(2) crystals at1.5% Yb doping,and the LSV and CV curves indicate that the Yb doping of 1.5% increases the oxygen precipitation potential and electrocatalytic oxidation activity of the electrode.When the electrode with Yb doping of 1.5% is the anode with current density of 10 mA/cm^(2) electrochemical oxidation time of 30 min,the electrode can remove chemical oxygen demand(COD) up to 85.06%,total organic carbon(TOC) up to 60.59% and UV_(254) from 1.594 to 0.507 for coking wastewater.Gas chromatography(GC-MS),UV-vis and three-dimensional fluorescence results of coking wastewater before and after treatment show that large toxic substances in coking wastewater are degraded to low toxic organic substances,and most soluble organic substances are degraded and transformed.This study provides the possibility of basic research for the engineering practice of electrochemical oxidation for the treatment of coking wastewater.

Treatment of coking wastewater by a novel electric assisted micro-electrolysis filter

A newly designed electric assisted micro-electrolysis filter（E-ME） was developed to investigate its degradation efficiency for coking wastewater and correlated characteristics. The performance of the E-ME system was compared with separate electrolysis（SE） and micro-electrolysis（ME） systems. The results showed a prominent synergistic effect on COD removal in E-ME systems. Gas chromatography/mass spectrometry（GC–MS） analysis confirmed that the applied electric field enhanced the degradation of phenolic compounds.Meanwhile, more biodegradable oxygen-bearing compounds were detected. SEM images of granular activated carbon（GAC） showed that inactivation and blocking were inhibited during the E-ME process. The effects of applied voltage and initial p H in E-ME systems were also studied. The best voltage value was 1 V, but synergistic effects existed even with lower applied voltage. E-ME systems exhibited some p H buffering capacity and attained the best efficiency in neutral media, which means that there is no need to adjust p H prior to or during the treatment process. Therefore, E-ME systems were confirmed as a promising technology for treatment of coking wastewater and other refractory wastewater.

Removal of total cyanide in coking wastewater during a coagulation process: Significance of organic polymers

Whether a cationic organic polymer can remove more total cyanide （TCN） than a non-ionic organic polymer during the same flocculation system has not been reported previously. In this study, the effects of organic polymers with different charge density on the removal mechanisms of TCN in coking wastewater are investigated by polyferric sulfate （PFS） with a cationic organic polymer （PFS-C） or a non-ionic polymer （PFS-N）. The coagulation experiments results show that residual concentrations of TCN （Fe（CN）6^3-） after PFS-C flocculation （TCN 〈 0.2 mg/L） are much lower than that after PFS-N precipitation. This can be attributed to the different TCN removal mechanisms of the individual organic polymers. To investigate the roles of organic polymers, physical and structural characteristics of the floes are analyzed by FT-IR, XPS, TEM and XRD. Owing to the presence of N＋ in PFS-C, Fe（CN）3- and negative flocs （Fe（CN）63- adsorbed on ferric hydroxides） can be removed via charge neutralization and electrostatic patch flocculation by the cationic organic polymer. However, non-ionic N in PFS-N barely reacts with cyanides through sweeping or bridging, which indicates that the non-ionic polymer has little influence on TCN removal.