Inhibition of zinc ions in sulfur-driven autotrophic denitrification process: What is the behavior of extracellular polymeric substances?

Sulfur-driven autotrophic denitrification (SAD) process is a cost-effective and sustainable method for nitrogen removal from wastewater. However, a higher concentration of zinc ions (Zn(II)) flowing into wastewater treatment plants poses a potential threat to the SAD process. This study examined that a half maximal inhibitory concentration (IC50) of Zn(II) was 7 mg·L-1 in the SAD process. Additionally, the addition of 20 mg·L-1 Zn(II) resulted in a severe accumulation of nitrite to 150.20 ± 6.00 mg·L-1 when the initial concentration of nitrate was 500 mg·L-1. Moreover, the activities of nitrate reductase, nitrite reductase, dehydrogenase and electron transport system were significantly inhibited under Zn(II) stress. The addition of Zn(II) inhibited EPS secretion and worsened electrochemical properties. The result was attributed to the spontaneous binding between EPS and Zn(II), with a ΔG of -17.50 KJ·mol-1 and a binding constant of 1.77 × 104 M-1, respectively. Meanwhile, the protein, fulvic acid, and humic-like substances occurred static quenching after Zn(II) addition, with -OH and -C=O groups providing binding sites. The binding sequence was fulvic acid→protein→humic acid and -OH â†’ -C=O. Zn(II) also reduced the content of α-helix, which was unfavorable for electron transfer. Additionally, the Zn(II) loosened protein structure, resulting in a 50 % decrease in α-helix/(ß-sheet+random coil). This study reveals the effect of Zn(II) on the SAD process and enhances our understanding of EPS behavior under metal ions stress.

Meta-analyzing the mechanism of pyrogenic biochar strengthens nitrogen removal performance in sulfur-driven autotrophic denitrification system: Evidence from metatranscriptomics.

Sulfur-driven autotrophic denitrification (SAD) exhibits significant benefits in treating low carbon/nitrogen wastewater. This study presents an eco-friendly, cost-effective, and highly efficient method for enhancing nitrogen removal performance. The addition of biochar prepared at 300 °C (BC300) notably increased nitrogen removal efficiency by 31.60 %. BC300 concurrently enhanced electron production, the activities of the electron transfer system, and electron acceptors. With BC300, the ratio of NADH/NAD+ rose 2.00±0.11 times compared to without biochar, and the expression of NAD(P)H dehydrogenase genes was markedly up-regulated. In the electron transfer system, BC300 improved the electroactivity of extracellular polymeric substances and the activities of NADH dehydrogenase and complex III in intracellular electron transfer. Subsequently, electrons were directed into denitrification enzymes, where the nar, nir, nor, and nos related genes were highly expressed with BC300 addition. Significantly, BC300 activated the Clp and quorum sensing systems, positively influencing numerous gene expressions and microbial communication. Furthermore, the O%, H%, molar O/C, and aromaticity index in biochar were identified as crucial bioavailable parameters for enhancing nitrogen removal in the SAD process. This study not only confirms the application potential of biochar in SAD, but also advances our comprehension of its underlying mechanisms.

Rapid start-up sulfur-driven autotrophic denitrification granular process: Extracellular electron transfer pathways and microbial community evolution.

Sulfur-driven autotrophic denitrification (SAD) granular process has significant advantages in treating low-carbon/nitrogen wastewater; however, the slow growth rate of sulfur-oxidizing bacteria (SOB) results in a prolonged start-up duration. In this study, the thiosulfate-driven autotrophic denitrification (TAD) was successfully initiated by inoculating anaerobic granular sludge on Day 7. Additionally, the electron donor was successfully transferred to the cheaper elemental sulfur from Day 32 to Day 54 at the nitrogen loading rate of 176.2 g N m-3 d-1. During long term experiment, the granules maintained compact structures with the α-helix/(ß-sheet + random coil) of 29.5-40.1 %. Extracellular electron transfer (EET) pathway shifted from indirect to direct when electron donors were switched thiosulfate to elemental sulfur. Microbial analysis suggested that thiosulfate improved EET involving enzymes activity. Thiobacillus and Sulfurimonas were dominant in TAD, whereas Longilinea was enriched in elemental sulfur-driven autotrophic denitrification. Overall, this strategy achieved in-situ enrichment of SOB in granules, thereby shortening start-up process.

[Prenatal ultrasonic manifestation and clinical significance of fetal hemivertebra].

OBJECTIVE: To explore the manifestation and clinical significance of prenatal ultrasound diagnosis of fetal hemivertebra. METHODS: In the study, 27 fetals with hemivertebras (proven by post-natal X-ray, CT or MR) were examined by prenatal ultrasound and MR in Women & Children's Hospital, Ningbo, Zhejiang. Two-dimensional and three-dimensional prenatal ultrasonic manifestations were retrospective analyzed and compared with the prenatal MR diagnoses, the post-natal X-ray, CT, MR examinations. All the fetuses were carried out karyotype examinations. The full-term births recieved surgical treatment and the Cobb angle correction rate was calculated. RESULTS: The 27 cases of fetal ultrasound showed the morphological changes of the spine, with the involved segment only half of the vertebra, in which 9 cases were single hemivertebra and 18 cases multiple, 8 cases were no malformation and 19 cases other malformations, and 19 cases were induction of labor, and 8 cases of term delivery. Compared with postpartum X-ray and other imaging tests, the prenatal ultrasound accuracy rate was 92.5%, and prenatal MR 96.3%. In the 27 cases, the chromosome cultures of 25 cases were successful, in which the normal karyotype was 68.0% (17/25), and abnormal karyotype 32.0% (8/25) with multiple hemivertebra accounting for 47.1% (8/17). In 8 cases with posterior approach of hemivertebra resection, 6 patients were less than and equal to 3 years old, whose average Cobb angle correction rate was 38.02%, and 2 more than 3 years old, whose average Cobb angle correction rate was 24.98%. CONCLUSION: Fetal hemivertebra have typical sonographic manifestations. In diagnosis of fetal hemivertebra, the accuracy of prenatal ultrasound is close to that of MR, which has important clinical implications in diagnosis and overall assessment of fetal hemivertebra, and can also provide appropriate clinical genetic reference.

[Modeling formation of aerobic granule and influence of hydrodynamic shear forces on granule diameter].

A one-dimension aerobic granule mathematical model was established, basing on mathematical biofilm model and activated sludge model. The model was used to simulate simple aerobic granule process such as nutrients removal, granule diameter evolution, cycle performance as well as depth profiles of DO and biomass. The effluent NH4(+) -N concentration decreased as the modeling processed. The simulation effluent NO3(-)-N concentration decreased to 3 mg x L(-1) as the granules grew. While the granule diameter increased from 1.1 mm on day 30 to 2.5 mm on day 100, the TN removal efficiency increased from less than 10% to 91%. The denitrification capacity was believed to enhance because the anoxic zone would be enlarged with the increasing granule diameter. The simultaneous nitrification and denitrification occurred inside the big aerobic granules. The oxygen permeating depth increased with the consumption of substrate. It was about 100-200 microm at the beginning of the aeration phase, and it turned to near 800 microm at the end of reaction. The autotrophs (AOB and NOB) were mostly located at the out layer where the DO concentration was high. The heterotrophic bacteria were distributed through the whole granule. As hydrodynamic shear coefficient k(de) increased from 0.25 (m x d)(-1) to 5 (m x d)(-1), the granule diameter under steady state decreased form 3.5 mm to 1.8 mm. The granule size under the dynamic steady-state decreased with the increasing hydrodynamic shear force. The granule size could be controlled by adjusting aeration intensity.

[Effect of aeration intensity on the nitrogen and phosphorus removal performance of AOA membrane bioreactors].

The ability of simultaneous phosphorus and nitrogen removal of sequencing batch membrane bioreactor run in anaerobic/oxic/ anoxic mode (AOA MBR) was examined under three aeration intensities [2.5, 3.75 and 5.0 m3 x (m2 x h)(-10]. The results showed that the averaged removals of COD were over 90% at different aeration intensities. And the higher aeration intensity was, the more ammonia nitrogen removal rate achieved. The removal rates of NH4(+) under the three aeration intensities were 84.7%, 90.6% and 93.8%, respectively. Total nitrogen removal rate increased with the increasing aeration intensity. But excessive aeration intensity reduced TN removal. The removal rates of TN under the three aeration intensities were 83.4%, 87.4% and 80.6%, respectively. Aeration intensity affected the denitrifying phosphorus ability of the AOA MBR. The ratio of denitrification phosphorus removal under the three aeration intensities were 20%, 30.2% and 26.7%, respectively.

[Preparation and characterization of polyvinyl alcohol microsphere pre-coated dynamic membranes].

To alleviate membrane fouling and reduce the price of membrane module, polyvinyl alcohol microsphere pre-coated dynamic membranes (PVA-MS/PCDMs) were investigated detailedly. It was prepared when the polyvinyl alcohol microsphere of the pre-coated reagent deposited on the surface and entered the pores of porous substrate common filter cloth which was low-cost by means of circulatory filtration. The morphology and structure of PVA-MS/PCDMs were examined using scanning electron microscope (SEM), and approximate maximal separated aperture, water contact angle, pure water permeation resistance and relative Zeta potential of the membrance surface were also determined. The results showed that pure water permeation resistance of PVA-MS/PCDMs was between 0.64 x 10(10) m(-1) and 3.84 x 10(10) m(-1) and the approximate maximal separated aperture reached microfiltration level. The hydrophilicity of PVA-MS/PCDMs increased with increasing PVA-MS pre-coated mass. The relative Zeta potential of the PVA-MS/PCDMs, which were measured using 0.01 mol x L(-1) KCl solution, pH 8.0 +/- 0.2 and at 20 degrees C, reached a peak value of -36.4 mV with pre-coated mass of 15.7 g x m(-2) PVA-MS on the membrance surface. In addition, evolutions of PVA-MS/PCDMs flux versus time were investigated using a 4 hours short-term filtration experiment at 9.5 kPa in an SMBR and the permeability coefficient of PVA-MS/PCDMs were measured by the extracellular polymeric substances (EPS) diffusion experiments. Results indicated that the PVA-MS/PCDMs presented dramatically high anti-fouling characteristics when the pre-coated mass of PVA-MS was 23.9-61.9 g x m(-2).

[Denitrifying phosphorus removal in MUCT-MBR].

The self-designed MUCT-MBR simplifies the MUCT process with reducing reactors from 5 to 2, which greatly reduces land occupied by equipment. Instead of secondary sedimentation tank, the membrane effluent quality is quite safe, and the operation is simple. In the investigation about simultaneous phosphorus and nitrogen removal of MUCT-MBR, the results showed that: when the proportions of C, N and P in the influent were 33.3/5/1-25/5.5/1, the average removal rate of COD, TN and TP in the whole experimental process were 89.3%, 75.4% and 79.2%, respectively. And the sludge settling capacity had no influence on the membrane effluent quality. The key factor of N and P removal rates is denitrifying phosphorus removal in anoxic condition. The proportion of denitrifying phosphate accumulating organisms (DPAOs) and the rate of denitrifying phosphorous removal were 84.2% and 67.07% on the 58th day, respectively.

[Combined MBR-RO process treating high strength wastewater].

The performances of A/O-MBR/RO system for the removal of nitrogen and COD were investigated. Result indicated that most organic was removed in the A/O-MBR and the average removal efficiency was 95.6%. The water quality of RO effluent which in terms of TOC < 0.9 mg x L(-1), TN < 12.65 mg x L(-1), total rigidity < 0.038 mol x L(-1), total alkalinity < 14.6 mg x L(-1) could meet the water quality requirements for the town wastewater reuse. The average removal efficiency of organic was almost unaffected by COD/N, but the process of TN removal was affected by COD/N. TN removal was primarily based on simultaneous nitrification and denitrification (SND) process occurred in the aerobic zone and the average removal efficiency of TN was 89.4% with average COD/N of 10.2. Both aerobic SND and conventional biological nitrification/denitrification contributed to nitrogen removal, the average removal efficiency of TN was 72%, 74% with average COD/N of 7.1 and 5.6. The fouling cake layer formed on the RO membrane surface was observed by scanning electric microscopy. The membrane fouling was characterized by Fourier transform infrared spectroscopy technique which showed that the major components of the foulants were soluble microbe products.

[Effects of aeration intensity on characteristics of aerobic granules in sequencing batch airlift reactor (SBAR)].

This study investigated the physical performances of mature aerobic granules in SBAR, including granular shape and structure, mean size and granular relative strength, and cell metabolic activities to excrete extracellular polymeric substances (EPS), and nitrification and microbial activity, which performed visible differences at superficial upflow velocity of 1.8 cm x s(-1) and 0.9 cm x s(-1) respectively. Results show that granular surface was more irregular at the low aeration intensity than that at the high aeration intensity and granular interstice performed increasing with it. In 55 days, comparing with high aeration intensity, the sludge productivity and the mean size of aerobic granules at the low aeration intensity were increased by 33% and 25%, respectively, while the granular relative strength and EPS were reduced by 6% and 12%, respectively. The sludge volume index in two reactors were about between 10 mL x g(-1) and 15 mL x g(-1), which implied that granules at both aerations behaved well settling ability. The two reactors still performed well nitrification characteristic and ability to remove COD. The activity of nitrifying bacteria in aerobic granules at the low aeration intensity was lower than that at the high aeration intensity, while heterotrophs performed higher activity at the low aeration intensity.

[Preparation and characterization of cross-linked polyvinyl alcohol microspheres as a novel precoating reagent in dynamic membrane].

In order to modify the membrane surface by adsorbing a precoated layer, cross-linked polyvinyl alcohol microspheres were prepared via an emulsion polymerization that PVA cross linked with glutaraldehyde which was extracted by anhydrous aether. Effects of concentration of PVA solution, the volume ratio of aqueous phase to oil phase, the selection of surface active agent (SAA) and its concentration on the stabilization of emulsion were studied. When the concentration of PVA solution was 2.7%, the volume ratio of PVA solution/n-heptane was 40:60 and the concentration of span-80 was 1.30 g/dL, the prepared microspheres contented our needs. The size of microspheres diminished with the stirring speed increasing under the condition of enough span-80. It became smaller with the accretion of an aimed cross-linking degree which has little affection on surface charge. The contact angle of PVA microspheres with distilled water showed that these microspheres were hydrophilic. Morphology of PVA microspheres were observed by SEM. The FTIR results suggested that functional groups varied with the change of aimed cross-linking degrees.

[Start-up of carbon tube membrane-aerated biofilm reactor for completely autotrophic nitrogen removal].

A laboratory-scale membrane-aerated biofilm bioreactor (MABR) equipped with nonwoven fabrics support around the gas-permeable carbon tube was developed for single-stage autotrophic nitrogen removal based on partial nitrification and anaerobic ammonium oxidization. Initially, the reactor was inoculated with nitrifying biomass. By decreasing the air pressure in the lumen of carbon tube step by step, We can obtain stable nitrite accumulation. Subsequently, the reactor was reinoculated with Anammox biomass to construct a quite stable biofilm system for completely autotrophic N removal. After 120 d of operation, 88.7% of NH4(+) -N removal rate, 48.65 mg/L of average TN in the effluent, 83.77% of maximal TN removal efficiency was achieved under the following conditions: air pressure of 0.015 MPa, HRT of 6 h, and influent NH4(+) -N of 200 mg/L +/- 10 mg/L.

[Reactivation characteristics of stored aerobic nitrifying granules].

A sequencing batch airlift reactor (SBAR) was conducted to study the performance of physical characteristics and respirometric activities during reactivation of aerobic granules after a 2-month storage time. Results showed that the color of the aerobic granules had completely become dark, the size and settleability did hardly change during the storage period. The color turned to be brownish-yellowish when the reactor was restarted for 2 weeks. The MLSS, granule size, and settleability increased rapidly along with reactivation. The heterotrophic activity resumed 86% within 1 day and was fully reactivated after 5 days. Then the COD removal rate was stabilized above 80%. Higher aeration rate and longer cycle time facilitated the reactivation of nitrifiers. When aeration rate was increased from 0.05 m3 x h(-1) to 0.10 m3 x h(-1) at 41st day, activities of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria were improved from 88% and 82% to 122% and 92%, respectively. The removal rate of ammonium-nitrogen promptly reached above 96%. Nitrite-oxidizing bacteria were fully reactivated after the cycle time was prolonged from 4 h to 6 h.

[Nitrification performance and microbial community analysis in carbon membrane-aerated biofilm reactor].

A carbon membrane-aerated biofilm reactor was developed to treat nitrogenous inorganic wastewater. Influent NH; -N concentrations and HRT were changed to investigate nitrification performance of reactor,oxygen utilization and NH4+ -N's removal loading. Biofilm's surface characteristics and dominant bacteria of nitrifier were analyzed. The results show that under the conditions of intra-membrane pressure of 0.017 MPa, influent NH4+ -N of 50 mg/L and HRT of 8 h NH4+ -N removal efficiency reaches 96% and effluent average nitrite is 17 mg/L, which benefits short-cut nitrification to a certain extent. The bacteria within biofilm consume all oxygen supplied through carbon membrane. The maximum specific removal rate of NH4+ -N is 9.7 g/(m2 x d), which is limited by the amount of bacteria grown onto carbon membrane's surface. Fluorescent in situ hybridization analysis indicates that within the biofilm Nitrosomonas and Nitrosospira are main ammonia-oxidizing bacteria and occupy about 19% and 21% of the total bacteria number, respectively. The Nitrobacter are not observed and Nitrospira are dominant nitrite-oxidizing bacteria, the fraction of which is 20% of total bacteria.

An effective way to select slow-growing nitrifying bacteria by providing a dynamic environment.

In this study, two laboratory scale sequencing batch reactors (SBRs) were conducted to study the stability of aerobic granules. The strategy was involved in stepwise increase in ammonium (NH4+-N) concentration in the influent. Results showed that the activity of nitrifying bacteria and diameter of the aerobic granules significantly increased with gradually increasing NH4+-N, which reached persistently new balances by homeostasis. As a result, the stability of aerobic granules was remarkably enhanced. The value of sludge volume index (SVI) was below 25 ml/g, the mean settling velocity was excellent up to 107 m/h. The NH4+-N removal efficiency averaged above 99% and total nitrogen (TN) removal was greatly enhanced and could reach 68%. Besides dissolved oxygen, the granules size was also a dominant factor to influence denitrification, which could gradually increase in variable conditions through homeostasis. Stable, dense and well-settling nitrifying granules can be developed for simultaneous nitrification and denitrification removal.

[Enrichment of denitrifying phosphate accumulating organisms in sequencing batch membrane bioreactors].

The enrichment and characteristics of denitrifying phosphate accumulating organisms (DPAO), which are capable of utilizing nitrate as electron acceptor, was investigated in a laboratory-scale sequencing batch membrane bioreactors (SBMBR). The results demonstrated that the proportion of DPAO increased from 19.4% to 69.6% of total phosphate accumulating organisms after anaerobic-aerobic and anaerobic-anoxic-aerobic phases. SBMBR system could operate steadily when 120 mg nitrate was added continuously during the anoxic phase every period. Simultaneous phosphate uptake and biological denitrification with good performance could be obtained in SBMBR operated in steady-state. Nitrate and phosphorus removal efficiency were above 100% and 84% respectively during anoxic phase, however, the effluent TP concentration was low than 0.5 mg/L, the total phosphorus removal efficiency was 96.1%. Furthermore, the ammonium nitrogen removal efficiency of SBMBR could be maintained at 92.2%, and the cumulation of nitrite and nitrate was not observed clearly.

[Anti-fouling characteristics of the novel precoating reagent in dynamic membrane bioreactor].

Further research was made on precoating reagents in dynamic membrane process, in which novel precoating reagent-polyvinyl alcohol microsphere (PVA-MS) was prepared through emulsive polymerization of PVA and glutaraldehyde (GA). Furthermore, polymerization mechanism and anti-fouling characteristics through adsorption of membrane major fouling substances EPS upon PVA-MS were studied. The results showed that hemiacetals reaction played a major role in emulsive polymerization process, as the quantity of hydroxyl on PVA was decreased a little, PVA-MS surface behaved good hydrophilic, and the adsorption of protein and amylose upon PVA microsphere was stable and low, which was 0.543 mg x g(-1) and 0.694 mg x g(-1) respectively. In addition, PVA-MS surface behaved electronic negativity, which acted electrostatic repulsion to active sludge floc. Upon this characteristics and data, it was concluded that membrane fouling was delayed in microscopy structure. Diameter of PVA-MS in precoating liquid was about 1.14 microm, and Zeta-potential of precoating liquid with different precoating reagent concentration was less than - 39 mV, which made PVA microsphere diffused and stable from each other, then sedimentate rapidly on porous support membrane surface and internal wall of hole path. Besides, the morphology of PVA-MS and dynamic membrane formed from PVA-MS on support membrane were observed through SEM.

[Effect of activated sludge properties on short-term membrane fouling in submerged membrane bioreactor based on statistical analysis].

The influence of activated sludge properties on membrane fouling was investigated using statistical method. The results show that extracelluler polymeric substances (EPS), soluble microbial products (SMP), suspended solids in the supernatant (SSs), dynamic viscosity (micro), relative hydrophobicity (RH) and Zeta potential all have a significant influence on membrane permeability during microfiltration of activated sludge wastewater. The pearson's correlation coefficient (r(p)) for linear correlations between membrane fouling resistance and these sludge properties are 0.898, 0.712, 0.810, 0.691, 0.837, -0.881, respectively. The statistical results also show that SMP, micro, SSs, Zeta potential, and RH are mostly determined by EPS, indicating that EPS is the main and essential factor affecting membrane fouling in activated sludge properties. The pearson's correlation coefficient (r(p)) for linear correlations between total EPS and other sludge properties are 0.682, 0.633, 0.783, -0.953, 0.877, respectively. EPS should be controlled in the operation of membrane bioreactor to achieve a suitable permeation.

[Experimental study on application of the boundary layer theory for determining steady aeration intensity of precoated dynamic membrane bioreactor].

The rheological behaviour of the low sludge concentration liquor in MBR was investigated and made a conclusion that this liquor approximated to the Newtonian fluid while the concentration of the sludge was less than 8000 mg x L(-1). Furthermore, when the laminar flow boundary layer thickness on the surface of flat membrane came up to the thickness of precoated dynamic membrane (PDM), the steady aeration intensity was calculated by using the boundary layer theory in the Newtonian hydrodynamics. In order to ensure the stability of the pre-coated dynamic membrane bioreactor (PDMBR), oxygen supply aeration intensity was chosen to supply the best dissolved oxygen (3-5 mg x L(-1) in the initial stages and gradually increased to the steady aeration intensity. The results indicated that this mode could enhance the stability of PDM. In the experiment period (31 d), effluent COD was less than 12.48 mg x L(-1) and its average removal rate was 97.49%, NH4+ -N was less than 5.27 mg x L(-1) and its average removal rate was 76.13%, while the operational pressure just increased to 27 kPa. During the last period of the experiment, the stability of the PDMBR was studied when the aeration intensity was more than the steady aeration intensity and it was found that the precoated layer had been brushed off from the surface of common filter cloth, so this phenomenon proved that using the boundary layer theory could determine steady aeration intensity of PDMBR.

[Comparisons of simultaneous phosphorus and nitrogen removal in sequencing batch membrane bioreactors].

Two SBMBRs run in AO and A2O mode were operated in parallel to compare their ability of simultaneous phosphorus and nitrogen removal. The results showed that the removals of COD and ammonium nitrogen were averaged over 90% and 95%, respectively. A2O MBR has the stronger anaerobic phosphorus release ability; its SPRR30 outdoes 47.5% compared to AO MBR. SPUR30 of A2O MBR was lower which may attribute to the higher effluent TP content. The ratio of DPAO was enhanced 57% compared to AO MBR. And when nitrate as the only electron accepter, the phosphorus uptake mass with unit electron transfer was 30% higher in A2O MBR. This two factors lead to the stronger denitrifying phosphorus removal ability of A2O MBR. Furthermore, the membrane fouling was mitigated in A2O MBR though the aerobic time was half to that of AO MBR. The membrane filter function made SBMBR's effluent free of the sludge settlement ability.