Effects of the internal recycling rate on biological nutrient removal and microbial community structure in a sequential anoxic/anaerobic membrane bioreactor

This study investigated the effects of the internal recycling rate on nutrients removal in a sequential anoxic/anaerobic membrane bioreactor (SAM). Microbial community structure in sludge from the SAM was studied using quinone profile method. Above 98% COD, 68% nitrogen, and 55% phosphorus removal efficiencies were achieved when the internal recycling rate was 2.5 times influent flow. At that rate, the optimum specific nitrate loading rate and COD/NO(3)-N ratio were found to be 2.24 mgNO(3)-N g(-1) MLSS h(-1) and 9.13, respectively. Batch tests demonstrated that anoxic condition suppressed phosphorus release, and that denitrification was also influenced by initial substrate concentration. Denitrification appeared to have some priority over phosphorus release for substrate uptake. Microbial community analysis revealed a predominance of the subclass beta-Proteobacteria. Furthermore, it was found that Rhodocyclus-related bacteria were efficient at phosphorus removal than Actinobacteria.     

Denitrification of wastewater containing high nitrate and calcium concentrations

The removal of nitrate from rinse wastewater generated in the stainless steel manufacturing process by denitrification in a sequential batch reactor (SBR) was studied. Two different inocula from wastewater treatment plants were tested. The use of an inoculum previously acclimated to high nitrate concentrations led to complete denitrification in 6h (denitrification rate: 22.8mg NO(3)(-)-N/gVSSh), using methanol as carbon source for a COD/N ratio of 4 and for a content of calcium in the wastewater of 150mg/L. Higher calcium concentrations led to a decrease in the biomass growth rate and in the denitrification rate. The optimum COD/N ratio was found to be 3.4, achieving 98% nitrate removal in 7h at a maximum rate of 30.4mg NO(3)(-)-N/gVSSh and very low residual COD in the effluent.     

Denitrification by thermophilic soil bacteria with ethanol as substrate in a USB reactor

Summary Thermophilic biological denitrification was studied in a laboratory-upflow sludge blanket reactor fed with ethanol as carbon and energy source. High denitrification efficiency (>98%) was obtained at an ethanol: nitrate ratio >2 and at a hydraulic retention time (HRT) of 5 hours. The performance of the system with respect to nitrate removal was very satisfactory (>95%), even at high nitrate (235 mg NO₃-N/L) and hydraulic (3 hours HRT) loading rates applied.A stable sludge was formed by spherical granules 1 to 3 mm in diameter with a content of 25,8 g VS/L and were almost exclusively composed of bacteria belonging to the genus Bacillus.     

The integration of methanogenesis with shortcut nitrification and denitrification in a combined UASB with MBR

A combined system consisting of an up-flow anaerobic sludge blanket (UASB) and an aerobic membrane bioreactor (MBR) was operated at 28-30 degrees C and pH 7.8-8.1 for the treatment of low-strength synthetic wastewater enriched with organic carbon and NH4Cl. The MBR slurry was recirculated into the UASB with a ratio of 50-800%. It was found that nitrite was able to accumulate steadily during the nitrification step in the MBR at a low TOC/NH4+-N ratio. The mixed liquid containing NOX(-)-N in the MBR was recirculated to the UASB, and denitrification rather than methanogenesis became the preferred pathway. Whereas, the less carbon requirement for denitrification via nitrite rather than nitrate allowed methanogenesis to proceed simultaneously in the same reactor. The combination of membrane filtration and partial nitrification in the MBR with simultaneous denitrification and methanogenesis in the UASB could stably reach 98% TOC removal and 48.1-82.8% TN removal with recirculation ratio increasing from 50% to 800%.     

Removal of highly elevated nitrate from drinking water by pH-heterogenized heterotrophic denitrification facilitated with ferrous sulfide-based autotrophic denitrification

The performance of acetic acid-supported pH-heterogenized heterotrophic denitrification (HD) facilitated with ferrous sulfide-based autotrophic denitrification (AD) was investigated in upflow activated carbon-packed column reactors for reliable removal of highly elevated nitrate (42 mg NO₃-N l⁻¹) in drinking water. The use of acetic acid as substrate provided sufficient internal carbon dioxide to completely eliminate the need of external pH adjustment for HD, but simultaneously created vertically heterogenized pH varying from 4.8 to 7.8 in the HD reactor. After 5-week acclimation, the HD reactor developed a moderate nitrate removal capacity with about one third of nitrate removal occurring in the acidic zone (pH 4.8–6.2). To increase the treatment reliability, acetic acid-supported HD was operated under 10% carbon limitation to remove >85% of nitrate, and ferrous sulfide-based AD was supplementally operated to remove residual nitrate and formed nitrite without excess of soluble organic carbon, nitrite or sulfate in the final effluent.     

以纸为碳源去除地下水硝酸盐的研究

研究了以纸为碳源和反应介质的生物反应器对水中硝酸盐的去除。结果表明,以纸为碳源的反应器启动快．反硝化反应受温度及水力停留时间影响大。25℃的反硝化速率是14℃的1．7倍。在室温25±1℃,进水硝酸盐氮浓度为45．2mg·L^-1、水力停留时间8．6h时,反应器对硝酸盐氮的去除率在99．6％以上,当水力停留时间为7．2h,氮去除率只有50％。反硝化反应受pH值和溶解氧的影响小,反应进行过程中,纸表面形成了生物膜,纸也被消耗了．采用反应器出水再经活性炭吸附的工艺流程处理高硝酸盐氮地下水,<33．9mg·L^-1的硝酸盐氮完全去除,没有出现NC2-N,最终出水水质DOC<11mg·L^-1。     

Nitrate depletion in the riparian zone of a small woodland stream

Field enrichments with nitrate in two spring-fed drainage lines within the riparian zone of a small woodland stream near Toronto, Ontario showed an absence of nitrate depletion. Laboratory experiments with riparian substrates overlain with nitrate enriched solutions revealed a loss of only 5–8% of the nitrate during 48 h incubation at 12°C. However, 22–24% of the initial nitrate was depleted between 24 and 48 h when a second set of substrate cores was incubated at 20°C. Short-term (3 h) incubations of fresh substrates amended with acetylene were used to estimate in situ denitrification potentials which varied from 0.05–3.19 g N g^–1 d^–1 for organic and sandy sediments. Denitrification potentials were highly correlated with initial nitrate content of substrate samples implying that low nitrate levels in ground water and riparian substrates may be an important factor in controlling denitrification rates. The efficiency of nitrate removal in spring-fed drainage lines is also limited by short water residence times of < 1 h within the riparian zone. These data suggest that routes of ground water movement and substrate characteristics are important in determining nitrate depletion within stream riparian areas.     

Simultaneous organics removal and bio-electrochemical denitrification in microbial fuel cells

Simultaneous organics removal and bio-electrochemical denitrification using a microbial fuel cell (MFC) reactor were investigated in this study. The electrons produced as a result of the microbial oxidation of glucose in the anodic chamber were transferred to the anode, which then flowed to the cathode in the cathodic chamber through a wire, where microorganisms used the transferred electrons to reduce the nitrate. The highest power output obtained on the MFCs was 1.7 mW/m(2) at a current density of 15 mA/m(2). The maximum volumetric nitrate removal rate was 0.084 mg NO(3)(-)-N cm(-2) (electrode surface area) day(-1). The coulombic efficiency was about 7%, which demonstrated that a substantial fraction of substrate was lost without current generation.     

Monitoring the denitrification of wastewater containing high concentrations of nitrate with methanol in a sulfur-packed reactor

Biological denitrification of high nitrate-containing wastewater was examined in a sulfur-packed column using a smaller amount of methanol than required stoichiometrically for heterotrophic denitrification. In the absence of methanol, the observed nitrate removal efficiency was only about 40%, and remained at 400 mg NO(3)(-)-N/l, which was due to an alkalinity deficiency of the pH buffer and of CO(2) as a carbon source. Complete denitrification was achieved by adding approximately 1.4 g methanol/g nitrate-nitrogen (NO(3)(-)-N) to a sulfur-packed reactor. As the methanol concentration increased, the overall nitrate removal efficiency increased. As influent methanol concentrations increased from 285 to 570, 855, and 1,140 mg/l, the value of Delta mg alkalinity as CaCO(3) consumed/Delta mg NO(3)(-)-N removed increased from -1.94 to -0.84, 0.24, and 0.96, and Delta mg SO(4)(2-) produced/Delta mg NO(3)(-)-N removed decreased from 4.42 to 3.57, 2.58, and 1.26, respectively. These results imply the co-occurrence of simultaneous autotrophic and heterotrophic denitrification. Sulfur-utilizing autotrophic denitrification in the presence of a small amount of methanol is very effective at decreasing both sulfate production and alkalinity consumption. Most of methanol added was removed completely in the effluent. A small amount of nitrite accumulated in the mixotrophic column, which was less than 20 mg NO(2)(-) -N/l, while under heterotrophic denitrification conditions, nitrite accumulated steadily and increased to 60 mg NO(2)(-) -N/l with increasing column height.     

基于响应面法对一株好氧反硝化菌脱氮效能优化

【目的】水体富营养化是当今我国水环境面临的重大水域环境问题,氮素超标排放是主要的引发因素之一。好氧反硝化菌构建同步硝化反硝化工艺比传统脱氮工艺优势更大。获得高效的好氧反硝化菌株并通过生长因子优化使脱氮效率达到最高。【方法】经过序批式生物反应器(Sequencing batch reactor,SBR)的定向驯化,筛选获得高效好氧反硝化菌株,采用响应面法优化好氧反硝化过程影响总氮去除效率的关键因子(碳氮、溶解氧、pH、温度)。【结果】从运行稳定的SBR反应器中定向筛选高效好氧反硝化菌株Pseudomonas T13,采用响应面法对碳氮比、pH和溶解氧关键因子综合优化获得在18 h内最高硝酸盐去除率95%,总氮去除率90%。该菌株的高效反硝化效果的适宜温度范围为25?30 °C;最适pH为中性偏碱;适宜的COD/NO3?-N为4:1以上;最佳溶解氧浓度在2.5 mg/L。【结论】从长期稳定运行的SBR反应器中筛选获得一株高效好氧反硝化菌Pseudomonas T13,硝酸盐还原酶比例占脱氮酶基因的30%以上,通过运行条件优化获得硝氮去除率达到90%以上,对强化废水脱氮工艺具有良好应用价值。     

关于好氧反硝化菌筛选方法的研究

采用污泥驯化手段富集好氧反硝化细菌,将得到的驯化污泥分离纯化,共得到105株菌。用测TN的方法对所筛菌株进行初筛,得到25株对TN去除率达到50％以上的菌株。用氮元素轨迹跟踪测定法复筛,证实这25株菌都可以在好氧条件下进行硝酸盐呼吸,其中24株菌的反硝化过程为：NO3^-N→NO2^-N→N2,研究中还发现在反硝化过程中硝酸盐和亚硝酸盐不存在明显竞争被利用的作用。同时还提出了可能实现短程同步硝化反硝化以及在反馈作用的调节下,加快硝化反应速度的观点。     

Substrate-dependent denitrification of abundant probe-defined denitrifying bacteria in activated sludge

The denitrification capacity of different phylogenetic bacterial groups was investigated on addition of different substrates in activated sludge from two nutrient-removal plants. Nitrate/nitrite consumption rates (CRs) were calculated from nitrate and nitrite biosensor, in situ measurements. The nitrate/nitrite CRs depended on the substrate added, and acetate alone or combined with other substrates yielded the highest rates (3-6 mg N gVSS(-1) h(-1)). The nitrate CRs were similar to the nitrite CRs for most substrates tested. The structure of the active denitrifying population was investigated using heterotrophic CO2 microautoradiography (HetCO2-MAR) and FISH. Probe-defined denitrifiers appeared as specialized substrate utilizers despite acetate being preferentially used by most of them. Azoarcus and Accumulibacter abundance in the two different sludges was related to differences in their substrate-specific nitrate/nitrite CRs. Aquaspirillum-related bacteria were the most abundant potential denitrifiers (c. 20% of biovolume); however, Accumulibacter (3-7%) and Azoarcus (2-13%) may have primarily driven denitrification by utilizing pyruvate, ethanol, and acetate. Activated sludge denitrification was potentially conducted by a diverse, versatile population including not only Betaproteobacteria (Aquaspirillum, Thauera, Accumulibacter, and Azoarcus) but also some Alphaproteobacteria and Gammaproteobacteria, as indicated by the assimilation of 14CO2 by these probe-defined groups with a complex substrate mixture as an electron donor and nitrite as an electron acceptor in HetCO2-MAR-FISH tests.     

Treatment of hydroponic wastewater by denitrification filters using plant prunings as the organic carbon source

This study investigated the feasibility of using pre-treated plant liquors as organic carbon sources for the treatment of hydroponic wastewater containing high nitrate-N (>300 mg N/L). The waste plant material was pre-treated to extract organic carbon-rich liquors. When this plant liquor was used as an organic carbon source in denitrification filters at the organic carbon:nitrogen dose rate of 3C:N, nitrate removal efficiencies were >95% and final effluent nitrate concentrations were consistently <20mg N/L. However, at this dose rate, relatively high concentrations (>140 mg/L) of organic carbon (fBOD5) remained in the final effluents. Therefore, a 'compromise' organic carbon:nitrogen dose rate (2C:N) was trialled, at which nitrate removal efficiencies were maintained at >85%, final effluent nitrate concentrations were consistently below 45 mg N/L, and effluent fBOD5 concentrations were <25mg/L. This study has demonstrated that waste plant material is a suitable carbon source for the removal of nitrate from hydroponic wastewater in a denitrification filter.     

Using the combined bioelectrochemical and sulfur autotrophic denitrification system for groundwater denitrification

A combined bioelectrochemical and sulfur autotrophic denitrification system (CBSAD) was evaluated to treat a groundwater with nitrate contamination (20.9-22.0mgNO(3)(-)-N/L). The reactor was operated continuously for several months with groundwater to maximize treatment efficiency under different hydraulic retention times (HRT) and electric currents. The denitrification rate of sulfur autotrophic part followed a half-order kinetics model. Moreover, the removal efficiency of bioelectrochemical part depended on the electric current. The reactor could be operated efficiently at the HRT ranged from 4.2 to 2.1h (corresponding nitrogen volume-loading rates varied from 0.12 to 0.24 kg N/m(3)d; and optimum current ranged from 30 to 1000 mA), and the NO(3)(-)-N removal rate ranged from 95% to 100% without NO(3)(-)-N accumulation. The pH of effluent was satisfactorily adjusted by bioelectrochemical part, and the sulfate concentration of effluent was lower than 250 mg/L, meeting the drinking water standard of China EPA.     

Kinetics of nitrification and denitrification reactions

Nitrification and denitrification are important microbiological reactions of nitrogen. In this work, the kinetics of these reactions have been investigated based on a Monod-type expression involving two growth limiting substrates: ammonium nitrogen and dissolved oxygen for nitrification and nitrate nitrogen and dissolved organic carbon for denitrification. The kinetic constants and yield coefficients were evaluated for both these reactions. Past experimental work was used to determine the constants for the nitrification reaction. For the denitrification reaction, experiments were performed in a stirred tank reactor under conditions such that only one substrate was growth limiting. Steady-state values of the substrate concentrations in the reactor were determined at various dilution rates. These data were analyzed to obtain the kinetic and stoichiometric constants. From these constants it was concluded that in the range of nitrate nitrogen concentrations encountered in waste water, the denitrification reaction can be considered a first-order reaction. It was also found that three times as much organic carbon is required as nitrate nitrogen for complete nitrogen removal.     

Nitrate removal with bacterial cells attached to quartz sand and zeolite from salty wastewaters

A mixed bacterial culture was acclimated to the removal of high nitrate-N concentrations (100–750 mg NO₃ ⁻-N L⁻¹) from salty wastewaters. The experiments were carried out under anoxic conditions in the presence of 0.5, 1.5 and 3% (w/v) NaCl at different temperatures. The acclimated mixed bacterial culture was attached to quartz sand and zeolite. Denitrification was monitored in a continuous-flow bioreactor at different hydraulic retention times (HRT). Nitrate removal with cells attached to quartz sand and zeolite was completed at HRT of 167 h and 25 h respectively. Then brine denitrification with bacterial cells attached to zeolite was monitored for 85 days. Under the increased nitrate loading rate, nitrate removal was above 90%. Furthermore, during denitrification, not more than 0.5 mg NO₂ ⁻-N L⁻¹ could be produced. It can be concluded that nitrate removal with the cells attached to zeolite is economically and operationally a promising solution to denitrification of brine wastewaters.     

Differential sensitivity of chicken progesterone receptor forms to sulfhydryl reactive reagents

DNA binding of chick progesterone receptor B form (PRB) has been examined and compared to that of the A form (PRA). We found that the elution profiles of the two receptors overlap on DNA-cellulose columns. Both PRA or PRB could bind to plasmid DNA equivalently as assayed by sedimentation velocity studies. However, DNA-binding activity of the two receptor forms showed differential sensitivity to reducing agents and to sulfhydryl (SH) reactive reagents. Reducing agents stabilized DNA-binding activity of PRA more efficiently than they stabilized PRB. Moreover, removal of reducing agents from receptor preparations caused preferential loss of DNA binding by PRB compared to the PRA. DNA-binding activity of PRA was readily destroyed by sulfhydryl modifying reagents such as N-ethylmaleimide and iodoacetamide while PRB was 3-4 times less sensitive to these reagents. We conclude the DNA-binding activity of PRB is less stable due to altered accessibility of SH groups despite the amino acid sequence identity of the DNA-binding domains of PRA and PRB.     

Simultaneous nitrification and denitrification in a mixed methanotrophic culture

Simultaneous nitrification and denitrification using a mixed methanotrophic culture was investigated. When both NO₃ ^–-N (108 mg l^–1) and NH₃-N (59 mg l^–1) were added into batch reactors, nitrate removal was complete within 10 h at the rate of 47 mg NO₃ ^–-N g VSS^–1 day^–1 when dissolved oxygen (DO) concentration was maintained at 2 mg DO l^–1. Ammonia removal started simultaneously with nitrate removal at a slower rate of 14 NH₃-N g VSS^–1 day^–1. No significant accumulation of nitrite or nitrate during ammonia utilization suggested the occurrence of simultaneous nitrification and denitrification.     

Nitrate removal in aquariums by immobilized pseudomonas

Biological denitrification of nitrate to nitrogen gas was examined in a freshwater and a marine aquarium. Nitrate removal in the aquarium water was accomplished with denitrifiers immobilized in a freeze-dried, alginate-starch matrix. Starch served as a bacterial carbon source and cellular matrix-strengthening filler. Freeze-dried beads were placed in canisters through which nitrate-rich aquarium water was recirculated. The freshwater aquarium (100 L) contained goldfish (Carassius auratus) at a total biomass of 390 g, whereas cichlids (Oreochromis mossambicus) were kept at a similar stocking density in the marine aquarium. Denitrification resulted in low ambient nitrate concentrations in both aquariums. The specific nitrate removal rate of the freshwater beads was significantly higher (50 microg of NO(3)-N/bead/day) than that of seawater beads (5 microg of NO(3)-N/bead/day). Differences in ambient nitrate concentrations between both aquariums and diffusion limitation of nitrate to the active denitrification sites within the beads might explain these observed differences.