WO2020057332A1 - 一种确定污水处理厌氧氨氧化生物膜最适保存温度的方法 - Google Patents

一种确定污水处理厌氧氨氧化生物膜最适保存温度的方法 Download PDF

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WO2020057332A1
WO2020057332A1 PCT/CN2019/102968 CN2019102968W WO2020057332A1 WO 2020057332 A1 WO2020057332 A1 WO 2020057332A1 CN 2019102968 W CN2019102968 W CN 2019102968W WO 2020057332 A1 WO2020057332 A1 WO 2020057332A1
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anaerobic ammonia
ammonia oxidation
biofilm
oxidation biofilm
buffer solution
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French (fr)
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王硕
朱引
衣雪松
李激
王燕
郑俊华
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江南大学
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Priority to US16/869,679 priority Critical patent/US11513053B2/en

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D71/00—Semi-permeable membranes for separation processes or apparatus characterised by the material; Manufacturing processes specially adapted therefor
    • B01D71/06—Organic material
    • B01D71/56—Polyamides, e.g. polyester-amides
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/02—Aerobic processes
    • C02F3/10—Packings; Fillings; Grids
    • C02F3/102—Permeable membranes
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/28—Anaerobic digestion processes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/04—Preserving or maintaining viable microorganisms
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/18—Water
    • G01N33/1826—Organic contamination in water
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00—Nature of the contaminant
    • C02F2101/10—Inorganic compounds
    • C02F2101/16—Nitrogen compounds, e.g. ammonia
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2209/00—Controlling or monitoring parameters in water treatment
    • C02F2209/02—Temperature
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2305/00—Use of specific compounds during water treatment
    • C02F2305/06—Nutrients for stimulating the growth of microorganisms
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/30—Aerobic and anaerobic processes
    • C02F3/302—Nitrification and denitrification treatment
    • C02F3/307—Nitrification and denitrification treatment characterised by direct conversion of nitrite to molecular nitrogen, e.g. by using the Anammox process
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N2015/1006—Investigating individual particles for cytology
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N2015/1488—Methods for deciding

Definitions

  • the invention relates to a method for determining the optimal storage temperature of an anaerobic ammonia oxidation biofilm for sewage treatment, and belongs to the technical field of environmental engineering.
  • Anaerobic ammonia oxidation technology refers to the process of anaerobic ammonia oxidizing bacteria using nitrite nitrogen to oxidize ammonia nitrogen under anaerobic conditions, and finally generate nitrogen.
  • anaerobic ammonia-oxidizing biofilm technology is used to implement a wastewater treatment plant based on efficient nitrogen removal and energy saving. Is of great significance.
  • the growth rate of anaerobic ammonia oxidizing bacteria is low, and the generation time is longer (about 15-20). d) If the anaerobic ammonia oxidizing bacteria can be attached to the suspended filler to form a biofilm, and then cultivated and preserved, it can effectively help the sewage treatment plant with a low carbon source and a tight land in a short time Start operation and make ammonia nitrogen and total nitrogen meet the standard discharge. Temperature is an important parameter that affects microbial activity.
  • Determining the temperature that is most suitable for anaerobic ammonia oxidation biofilm storage will help simplify the microbial activity recovery process of anaerobic ammonia oxidation biofilm and shorten the start of engineering applications Time to achieve the effect of energy saving.
  • the determination of the optimal storage temperature by the existing method requires re-inoculation of the anaerobic ammonia oxidation biofilm in the bioreactor, which takes about 8-35. Only then can the activity recovery effect of the anaerobic ammonia oxidation biofilm be determined, which becomes the key to restrict the engineering application of the anaerobic ammonia oxidation biofilm.
  • the ammonia nitrogen and total nitrogen indicators of the sewage treatment plant will be discharged in a short time, and at the same time, the effect of land saving and energy saving will be achieved.
  • the present invention is based on the flow cytometry to characterize the cell activity state of the anaerobic ammonia oxidation biofilm preserved under different temperature conditions.
  • a first object of the present invention is to provide a method for confirming the optimal storage temperature of an anaerobic ammonia oxidation biofilm for sewage treatment.
  • the method is based on measuring the cell activity state of the anaerobic ammonia oxidation biofilm based on flow cytometry. Compare the measurement results of the anaerobic ammonia-oxidized biofilms stored at different temperatures with the anaerobic ammonia-oxidized biofilms before storage. Storage temperature.
  • the determination of the cell activity state of the anaerobic ammonia-oxidizing biofilm includes the content determination of live cells, early apoptotic cells, late apoptotic cells, and dead cells.
  • One aspect of the present invention is an embodiment, and the step of confirming an optimal temperature by flow cytometry includes:
  • the buffer includes a phosphate buffer and fetal bovine serum.
  • the phosphate buffer solution includes 0.2 mol / L disodium hydrogen phosphate and 0.2 mol / L sodium dihydrogen phosphate.
  • One aspect of the present invention is an embodiment, and a volume ratio of the phosphate buffer solution and fetal bovine serum in the buffer solution is 8: 1 to 10: 1.
  • One embodiment of the present invention is that the pH of the buffer solution is 7.2 to 8.0, and the dilution ratio with the anaerobic ammonia oxidation biofilm is 8 to 10: 1.
  • One aspect of the present invention is an embodiment, and the filtering is selected from 6 to 8 ⁇ m pore size nylon membrane.
  • the centrifugal speed is 5000 ⁇ 10000 rpm / min.
  • the sample serum and 10x The mixing volume ratio of Annexin V Binding Buffer is 1: 2 ⁇ 4.
  • the flow cytometer is used to determine the cell activity status of each sample in the control FITC.
  • 0.5 ⁇ l PI stain was added to Annexin V group, 0.5 was added to control PI group ⁇ l FITC Annexin V, 0.5 ⁇ l FITC Annexin V and 0.5 were added to the test group ⁇ l PI, mix and incubate in the dark at room temperature, and then test it on a flow cytometer.
  • the incubation is 10-20 min.
  • a second object of the present invention is to provide a method for quickly starting anaerobic ammonia oxidation biofilm wastewater treatment.
  • the method is to mature the anaerobic ammonia oxidation biofilm in advance and place it in a storage substrate at the optimal storage temperature. After preservation, the activity can be used for sewage treatment after recovery; the optimum temperature is confirmed by the above method.
  • the storage matrix includes: KHCO 3 1500 mg / L, K 2 HPO 4 15 mg / L, MgSO 4 180 mg / L, CaCl 2 20 mg / L; storage matrix NH 4 + - The N concentration was 50 mg / L, and the NO 2 -- N concentration was 75 mg / L.
  • One embodiment of the present invention is to restore the activity by inoculating an anaerobic ammonia-oxidizing biofilm into a bioreactor.
  • the bioreactor is pre-passed with N 2 to reduce the inhibition of anaerobic ammonia-oxidizing bacteria by oxygen.
  • the HRT is set to 4 h, the filling ratio of anaerobic ammonia oxidation biofilm suspension filler is 40%.
  • a third object of the present invention is to apply the method to sewage treatment.
  • the present invention uses flow cytometry to characterize the ratio of living cells, early apoptotic cells, late apoptotic cells and dead cells in anaerobic ammonia-oxidizing biofilms, confirming the optimal storage temperature within a few hours, and comparing with Correlation analysis was performed on the characteristic indexes of the recovery process of membrane activity, and a method for determining the optimal storage temperature of anaerobic ammonia oxidation biofilm based on flow cytometry was established.
  • the step of recovering the activity of the suspended filler biofilm can be omitted, which can effectively help the sewage treatment plant that prepares to use the anaerobic ammonia oxidation biofilm technology to achieve ammonia nitrogen and total nitrogen discharge standards to achieve land saving and energy saving operations, and can effectively shorten
  • the start-up time of the anaerobic ammonia oxidation biofilm engineering application to maintain the long-term stable operation of the anaerobic ammonia oxidation biofilm process is highly feasible.
  • Figure 1 shows the change of extracellular polymer PN / PS
  • Figure 2 is the removal rate of ammonia nitrogen (AN);
  • FIG. 3 shows the removal rate of total nitrogen (TN).
  • the sewage of the sewage treatment plant of the present invention includes domestic water in upstream residential areas and a small part of industrial wastewater, and the average annual water intake is COD 236mg / L, ammonia nitrogen 30.1 mg / L, total nitrogen 37.8 mg / L, total phosphorus 4.5 mg / L, nitrate nitrogen content is less than 1.0 mg / L.
  • the storage temperature of anaerobic ammonia oxidation biofilm was set to -20 ° C, 4 ° C and 20 ° C.
  • 150 anaerobic ammonia oxidation biofilm suspension fillers in the sludge dewatering filtrate treatment tank of the sewage treatment plant were taken out, and an average of three equal portions were placed in 1000 ml serum bottles containing a 750 ml storage matrix (the serum bottles were pre-filled with N 2 To remove O 2 from the air, the storage matrix provides NH 4 + -N and NO 2 -- N with NH 4 Cl and NaNO 2.
  • the anaerobic ammonia-oxidizing biofilm After being stored at -20 ° C, 4 ° C, and 20 ° C for more than 120 days, the anaerobic ammonia-oxidizing biofilm is used to determine the cell status of the anaerobic ammonia-oxidizing biofilm.
  • the specific flow cytometry cell status test steps are as follows:
  • the test results of the anaerobic ammonia oxidation biofilm cells are shown in Table 1.
  • the proportion of living cells in the anaerobic ammonia oxidation biofilm in the sewage treatment plant sludge dewatering filtrate treatment tank is relatively high, indicating that the sewage treatment plant sludge dehydration filtrate treats high ammonia nitrogen.
  • the content of living cells stored at 20 ° C for the determination of anaerobic ammonia oxidation biofilm is the lowest, indicating that 20 ° C is not suitable for the preservation of anaerobic ammonia oxidation biofilm.
  • the anaerobic ammonia oxidation biofilm stored at 4 °C has the highest proportion of living cells, reaching 65.0%, and the proportion of late apoptotic cells and dead cells is about 17.8%, which proves that 4 °C conditions can be used for the preservation of anaerobic ammonia oxidation biofilm.
  • the proportion of living cells of anaerobic ammonia oxidation biofilm reaches 50.1%, which is only 22.9% lower than the proportion of living cells of anaerobic ammonia oxidation biofilm stored at 4 °C.
  • the ratio of cells to dead cells is about 39.5%, indicating that storage conditions at -20 ° C are not suitable for the preservation of anaerobic ammonia-oxidized biofilms. Therefore, 4 ° C was preliminarily determined as the optimal temperature for storing anaerobic ammonia oxidation biofilm.
  • Sequential batch reactor is selected.
  • the specific operating conditions include: taking anaerobic ammonia oxidation biofilms from different serum bottles and inoculating them in a bioreactor (effective volume 10.0 L) to activate the anaerobic ammonia oxidation biofilms. restore.
  • the anaerobic ammonia oxidation biofilms stored at -20 ° C, 4 ° C and 20 ° C were placed in R1, R2 and R3, respectively.
  • the bioreactor was pre-passed with N 2 to reduce the inhibition of anaerobic ammonia-oxidizing bacteria by oxygen.
  • the HRT was set to 4 h, and the filling ratio of the anaerobic ammonia-oxidizing biofilm suspension filler was 40%.
  • anaerobic ammonia oxidation biofilms in R1, R2, and R3 are shown in Table 2. After the activity of anaerobic ammonia oxidation biofilms was restored, they were stored at 4 ° C and -20 ° C. The density and thickness of the oxidized ammonia-oxidized biofilm were close to those of the anaerobic ammonia-oxidized biofilm before storage, and only the density and thickness of the anaerobic ammonia-oxidized biofilm stored at 20 ° C decreased slightly.
  • the biomass of anaerobic ammonia-oxidized biofilms has decreased, but after recovery of activity, the biomass of anaerobic ammonia-oxidized biofilms stored at 4 ° C and -20 ° C is close to the anaerobic ammonia before storage.
  • the oxidized biofilm biomass indicates that the anaerobic ammonia-oxidized biofilm re-adapted to the environment and the biomass increased steadily.
  • the anaerobic ammonia oxidation biofilm has 0.27 gN / gMLVSS • d and 2.3 umol / gVSS respectively than the anaerobic ammonia oxidation activity and heme c content.
  • the anaerobic ammonia oxidation biofilm domesticated by the sewage treatment plant reaches the same specific anaerobic ammonia oxidation. Active and hemoglobin c contents took 32 and 24 days, respectively.
  • the time required for the anaerobic ammonia oxidation biofilm in R1 to reach the same specific anaerobic ammonia oxidation activity and heme c content takes 9 and 13 days, respectively, and the anaerobic ammonia oxidation in R2 It takes 7 and 11 days for the biofilm to reach the same specific anaerobic ammonia oxidation activity and heme c content, respectively.
  • the time for the anaerobic ammonia oxidation biofilm in R3 to reach the same specific anaerobic ammonia oxidation activity and heme c content takes 11 respectively. And 17 d. This shows that the anaerobic ammonia-oxidized biofilms after recovery of activity have better denitrification effect. Among them, the anaerobic ammonia-oxidized biofilms stored at 4 ° C have the shortest recovery time of microbial activity and are more suitable for storing anaerobic ammonia-oxidized organisms. membrane.
  • Extracellular polymer is an important factor in the formation of anaerobic ammonia oxidation biofilms, and the ratio of polysaccharides (PS) to protein (PN) in extracellular polymers (PS / PN) is a measure of anaerobic ammonia oxidation Important parameters for membrane structure stability.
  • PS / PN polysaccharides
  • PN protein
  • PS / PN in anaerobic ammonia oxidation biofilm in R1 shows a trend of first decrease and then increase, indicating that the stability of anaerobic ammonia oxidation biofilm stored at -20 °C is gradually restored.
  • the anaerobic ammonia oxidation biofilm PS / PN in R3 was gradually increased and then gradually decreased and finally stabilized, indicating that 20 ° C is not suitable for storing anaerobic ammonia oxidation biofilm
  • R2 anaerobic ammonia oxidation biofilm PS / PN
  • the overall fluctuation is small, which indicates that the anaerobic ammonia-oxidized biofilm stored at 4 °C maintains high stability after its activity is restored.
  • 4 °C is suitable as the storage temperature of anaerobic ammonia-oxidized biofilm.
  • the fastest recovery of the anaerobic ammonia oxidation biofilm in R2 is higher than the anaerobic ammonia oxidation activity corresponding to the heme c content, indicating that the condition of 4 ° C is more suitable for the preservation of the anaerobic ammonia oxidation biofilm. Feasibility.
  • the anaerobic ammonia oxidation biofilm cell state was analyzed by flow cytometry. The results are shown in Table 3.
  • the content of living cells in the anaerobic ammonia oxidation biofilm at different storage temperatures is basically the same as the content of living cells in the anaerobic ammonia oxidation biofilm of the sewage treatment plant, which indicates that after the recovery of activity, the anaerobic ammonia oxidation biofilm can play a stable content. Nitrogen pollutant removal effect.
  • anaerobic ammonia-oxidized biofilm cells have the highest activity, which is more suitable as a condition for anaerobic ammonia-oxidized biofilms.
  • the anaerobic ammonia oxidation biofilm has a higher correlation than the anaerobic ammonia oxidation activity and heme c content and the proportion of living cells in the anaerobic ammonia oxidation biofilm.
  • the correlation coefficients are respectively It is as high as 0.9974 and 0.9930, indicating that it is extremely feasible to use the ratio of anaerobic ammonia-oxidized biofilm to viable cells as a method to evaluate the activity of anaerobic ammonia-oxidized biofilm.
  • the proportion of viable cells in the anaerobic ammonia-oxidized biofilms was the highest under the storage condition of 4 ° C, which was consistent with the results of the ratio of viable cells in anaerobic ammonia-oxidized biofilms after R2 was restored.
  • 4 ° C is the optimal temperature for storing anaerobic ammonia oxidation biofilm
  • flow cytometry can be used as a basis for determining the optimal storage temperature for anaerobic ammonia oxidation biofilm.
  • Flow cytometry is easy to operate, the data is fast, easy to obtain, accurate and reliable. It can also omit the anaerobic ammonia oxidation biofilm activity recovery process, which is of great significance for the preservation and recovery of anaerobic ammonia oxidation biofilm.
  • the storage temperature of anaerobic ammonia oxidation biofilm was set to -20 ° C, 4 ° C and 20 ° C.
  • 150 anaerobic ammonia oxidation biofilm suspension fillers in the sludge dewatering filtrate treatment tank of the sewage treatment plant were taken out, and an average of three equal portions were placed in 1000 ml serum bottles containing a 750 ml storage matrix (the serum bottles were pre-filled with N 2 To remove O 2 from the air, the storage matrix provides NH 4 + -N and NO 2 -- N with NH 4 Cl and NaNO 2.
  • the anaerobic ammonia-oxidizing biofilm After being stored at -20 ° C, 4 ° C, and 20 ° C for more than 120 days, the anaerobic ammonia-oxidizing biofilm is used to determine the cell status of the anaerobic ammonia-oxidizing biofilm.
  • the specific flow cytometry cell status test steps are as follows:
  • the inventors also found that the pH of the buffer has a greater impact on the state of the cells during the test, and the analysis of cell vitality data has a weak alkaline pH 7.2 ⁇ 8.0 environment, neutral partial acid or alkaline There is a small gap in cell status data under large conditions, and results cannot be accurately judged.
  • the inventors also examined the samples prepared with 6 ⁇ m and 10 ⁇ m pore diameters.
  • the test results of 6 ⁇ m samples are consistent with the verification experiments and the data is reliable.
  • the corresponding data of 10 ⁇ m does not have the analytical power and cannot be used to determine the most suitable storage temperature.

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Abstract

一种确定污水处理厌氧氨氧化生物膜最适保存温度的方法，其通过流式细胞术对厌氧氨氧化生物膜中活细胞、凋亡早期细胞、凋亡晚期细胞和死细胞的含量测定，从而测得最适保存温度。还公开了一种快速启动厌氧氨氧化生物膜污水处理的方法及相应方法在污水处理中的应用。

Description

一种确定污水处理厌氧氨氧化生物膜最适保存温度的方法 技术领域
本发明涉及一种确定污水处理厌氧氨氧化生物膜最适保存温度的方法，属于环境工程技术领域。
 背景技术
污水处理厂进水有机物含量偏低一直是制约总氮达标排放的技术难点，同时随着污水处理排放标准的提高，大量土地被改扩建工程占用，大量有机碳源被投加到缺氧池中，显著增加了投资建设和运营成本，进而严重影响污水处理厂的节能降耗效果。因此，基于节地目标和低碳源利用的污水脱氮技术被日益重视。厌氧氨氧化技术是指在厌氧条件下，厌氧氨氧化菌利用亚硝态氮氧化氨氮，最终生成氮气的过程。此工艺无需额外投加碳源，且厌氧氨氧化菌可以较好的附着在悬浮填料上，因此，厌氧氨氧化生物膜法技术对实现基于高效脱氮和节能降耗的污水处理厂运行具有重要意义。
但是在工程实例中厌氧氨氧化菌生长速率较低，代时较长（约15-20 d），如果可以将厌氧氨氧化菌附着在悬浮填料上以形成生物膜，进而培养成熟并予以保存，则可以有效帮助进水碳源较低，土地紧张的污水处理厂在较短时间内启动运行，并使氨氮和总氮达标排放。温度是影响微生物活性的重要参数，确定最适于厌氧氨氧化生物膜保存的温度，有助于简化厌氧氨氧化生物膜的微生物活性恢复过程，缩短厌氧氨氧化生物膜工程化应用启动时间，实现节能降耗效果。但现有方法对于最适保存温度的确定需要将厌氧氨氧化生物膜重新接种于生物反应器中，约需8-35 d才能确定厌氧氨氧化生物膜的活性恢复效果，成为制约厌氧氨氧化生物膜工程化应用的关键。
发明内容
为了简化厌氧氨氧化生物膜微生物活性的恢复过程，使污水处理厂氨氮和总氮指标短时间内达标排放，同时实现节地和节能降耗效果。本发明基于流式细胞术对不同温度条件下保存的厌氧氨氧化生物膜的细胞活性状态进行表征，通过厌氧氨氧化生物膜活性恢复效果和污泥微生物活性恢复后细胞活性状态，对流式细胞术的表征结果进行验证，最终建立基于流式细胞术的确定厌氧氨氧化生物膜最适保存温度的方法，为污水处理厂高标准污染物排放与节能降耗运行提供技术支撑。
本发明的第一个目的是提供一种确认污水处理厌氧氨氧化生物膜最适保存温度的方法，所述方法是基于流式细胞术对厌氧氨氧化生物膜的细胞活性状态进行测定，对比不同温度保存的厌氧氨氧化生物膜与保存前厌氧氨氧化生物膜的细胞活性状态的测定结果，以最接近保存前厌氧氨氧化生物膜的细胞活性状态时保存的温度作为最适保存温度。
本发明的一个是实施例，所述厌氧氨氧化生物膜的细胞活性状态测定包括活细胞、凋亡早期细胞、凋亡晚期细胞和死细胞的含量测定。
本发明的一个是实施例，所述流式细胞术确认最适温度的步骤包括：
（1）厌氧氨氧化生物膜测试样液的制备：取污泥脱水滤液处理池中的厌氧氨氧化生物膜，用缓冲液稀释厌氧氨氧化生物膜样品，震荡均匀后，过滤、离心，留用上清液，使用预冷磷酸盐缓冲液吹洗细胞，重复离心和清洗两次，再取上清液作为样品，使用适量10x Annexin V Binding Buffer混匀制得；
（2）置于流式细胞仪测定各样液的细胞活性状态。
本发明的一个是实施例，所述缓冲液包括磷酸盐缓冲液和胎牛血清。
本发明的一个是实施例，所述磷酸盐缓冲液包括0.2 mol/L磷酸氢二钠和0.2 mol/L磷酸二氢钠。
本发明的一个是实施例，所述缓冲液中的磷酸盐缓冲液和胎牛血清的体积比为8：1~10：1。
本发明的一个是实施例，所述缓冲液的pH为7.2~8.0，与厌氧氨氧化生物膜的稀释比为8~10：1。
本发明的一个是实施例，所述过滤是选用6~8 μm孔径的尼龙膜。
在本发明的一种实施方式中，所述离心速度为5000~10000 rpm/min。
在本发明的一种实施方式中，所述样品清液与10x Annexin V Binding Buffer的混合体积比例为1：2~4。
在本发明的一种实施方式中，所述流式细胞仪测定各样液的细胞活性状态是在对照FITC Annexin V组加入0.5μl PI染色剂，对照PI组加入0.5 μl FITC Annexin V，检测组加入0.5μl FITC Annexin V和0.5 μl PI，混匀后室温下避光孵育，之后于流式细胞仪上机检测。
在本发明的一种实施方式中，所述孵育为10-20 min。
本发明的第二个目的是提供一种快速启动厌氧氨氧化生物膜污水处理的方法，所述方法是预先将厌氧氨氧化生物膜培养成熟，置于保存基质中在最适保存温度下进行保存，活性恢复后即可用于污水处理；所述最适温度是利用上述方法确认。
本发明的一个是实施例，所述保存基质包括：KHCO 3 1500 mg/L, K 2HPO 4 15 mg/L，MgSO 4 180 mg/L，CaCl 2 20 mg/L；保存基质NH 4 +-N浓度为50 mg/L，NO 2 --N浓度为75 mg/L。
本发明的一个是实施例，所述活性恢复是将厌氧氨氧化生物膜接种于生物反应器中，生物反应器预先通N 2以降低氧气对厌氧氨氧化菌的抑制，HRT设置为4 h，厌氧氨氧化生物膜悬浮填料填充比为40%。
本发明的第三个目的是将所述方法应用在污水处理中的。
本发明有益效果
本发明通过流式细胞术对厌氧氨氧化生物膜中活细胞，凋亡早期细胞，凋亡晚期细胞和死细胞比例进行表征，数小时内确认最适保存温度，并与厌氧氨氧化生物膜活性恢复过程的特征指标进行关联性分析，建立了基于流式细胞术的确定厌氧氨氧化生物膜最适保存温度的方法。运用此方法，可以省略悬浮填料生物膜活性恢复的步骤，有效帮助准备采用厌氧氨氧化生物膜技术进行氨氮和总氮达标排放的污水处理厂实现节地和节能降耗运行，同时可以有效缩短厌氧氨氧化生物膜工程化应用启动时间，维持厌氧氨氧化生物膜工艺长期稳定运行，具有很高的可行性。
 附图说明
图1为胞外聚合物PN/PS的变化；
图2为氨氮（AN）的去除率；
图3为总氮（TN）的去除率。
具体实施方式
本发明污水处理厂污水包括上游各居民区的生活用水和少部分工业废水，进水年平均值为COD 236mg/L，氨氮30.1 mg/L，总氮37.8 mg/L，总磷4.5 mg/L，硝态氮含量低于1.0 mg/L。
实施例1
厌氧氨氧化生物膜保存培养：
厌氧氨氧化生物膜的保存温度设置为-20℃，4℃和20℃。将污水处理厂污泥脱水滤液处理池中的厌氧氨氧化生物膜悬浮填料150个取出，平均三等份分别置于装有750 ml保存基质的1000 ml血清瓶中（血清瓶预先充N 2以排出空气中的O 2），保存基质以NH 4Cl和NaNO 2提供NH 4 +-N和NO 2 --N，其余成分如下所示： KHCO 3 1500 mg/L, K 2HPO 4 15 mg/L，MgSO 4 180 mg/L，CaCl 2 20 mg/L；保存基质NH 4 +-N浓度为50 mg/L，NO 2 --N浓度为75 mg/L。将血清瓶（每个保存温度下设置3个平行样）分别置于-20℃，4℃和20℃，静止遮光保存。
保存的 厌氧氨氧化 生物膜细胞状态测试：
于-20℃，4℃和20℃保存的厌氧氨氧化生物膜在保存超过120 d后，用于测定厌氧氨氧化生物膜细胞状态，具体的流式细胞术细胞状态测试步骤如下：
（1）取10 ml厌氧氨氧化生物膜，用pH为7.8的磷酸盐缓冲液/胎牛血清的混合缓冲液稀释至100 ml，于涡旋仪漩涡震荡2 min，使生物膜破碎为絮体并保证均匀分布；
（2）用8μm孔径的尼龙膜过滤破碎后的样品，取1.5 ml置于1.5 ml尖底离心管中；
（3）样品于8000 rpm/min 离心5 min；
（4）用移液枪吸取离心后的样品上清液，留下约0.1 ml样品，使用预冷磷酸盐缓冲液/胎牛血清的混合缓冲液（pH为7.8）吹洗细胞，重复离心和清洗两次；
（5）离心后的样品用移液枪吸取上清液，留下约0.1 ml样品，使用0.3 ml 10x Annexin V Binding Buffer混匀；
（6）对照FITC Annexin V组加入0.5μl PI染色剂，对照PI组加入0.5 μl FITC Annexin V，检测组加入0.5μl FITC Annexin V和0.5 μl PI，混匀后室温下避光孵育15 min，之后于流式细胞仪上机检测。
厌氧氨氧化生物膜细胞状态测试结果如表1所示，污水处理厂污泥脱水滤液处理池中厌氧氨氧化生物膜活细胞比例较高，表明污水处理厂污泥脱水滤液对高氨氮处理效果良好。保存于20℃的用于测定厌氧氨氧化生物膜的活细胞含量最低，表明20℃不适于厌氧氨氧化生物膜的保存。保存于4℃的厌氧氨氧化生物膜其活细胞比例最高，达到65.0%，凋亡晚期细胞和死细胞比例约为17.8%，证明4℃条件可以用于厌氧氨氧化生物膜的保存。保存温度为-20℃时，厌氧氨氧化生物膜活细胞比例达到50.1%，仅比4℃保存的厌氧氨氧化生物膜活细胞比例低22.9%，但厌氧氨氧化生物膜凋亡晚期细胞和死细胞比例约为39.5%，表明-20℃的保存条件亦不适于保存厌氧氨氧化生物膜。因此，初步确定4℃为保存厌氧氨氧化生物膜的最适温度。
表1 保存120 d后的厌氧氨氧化生物膜细胞状态
厌氧氨氧化生物膜 活细胞 凋亡早期细胞 凋亡晚期细胞 死细胞
污泥脱水滤液处理池 82.5±4.8 3.7±1.6 5.9±2.1 7.9±1.6
保存于-20℃ 50.1±3.0 20.8±1.6 18.7±1.9 10.4±1.9
保存于4℃ 65.0±3.5 17.2±1.9 15.5±2.1 2.3±0.2
保存于20℃ 35.8±3.5 28.6±3.1 27.9±3.0 7.7±0.8
实施例2保存的厌氧氨氧化生物膜的活性恢复
选用序批式反应器，具体运行条件包括：取源于不同血清瓶的厌氧氨氧化生物膜，接种于生物反应器（有效容积10.0 L）中，用以对厌氧氨氧化生物膜进行活性恢复。于-20℃，4℃和20℃保存的厌氧氨氧化生物膜分别置于R1，R2和R3中。生物反应器预先通N 2以降低氧气对厌氧氨氧化菌的抑制，HRT设置为4 h，厌氧氨氧化生物膜悬浮填料填充比为40%。
实施例3活性恢复后厌氧氨氧化生物膜特性
经过30 d的活性恢复后，R1，R2和R3中的厌氧氨氧化生物膜特性如表2所示，在厌氧氨氧化生物膜活性恢复后，保存于4℃和-20℃温度下厌氧氨氧化生物膜密度和厚度与保存之前的厌氧氨氧化生物膜较为接近，只有保存在20℃的厌氧氨氧化生物膜密度和厚度略有下降。不同保存温度下厌氧氨氧化生物膜的生物量均有所降低，但经过活性恢复后，4℃和-20℃温度下保存的厌氧氨氧化生物膜的生物量已接近保存前厌氧氨氧化生物膜的生物量，说明厌氧氨氧化生物膜重新适应环境，生物量稳定增加。通常厌氧氨氧化生物膜比厌氧氨氧化活性和血红素c含量分别为0.27 gN/gMLVSS•d和2.3 umol/gVSS，污水处理厂驯化的厌氧氨氧化生物膜达到相同比厌氧氨氧化活性和血红素c含量的时间分别需要32和24 d。将保存的厌氧氨氧化生物膜进行活性恢复后，R1中厌氧氨氧化生物膜达到相同比厌氧氨氧化活性和血红素c含量的时间分别需要9和13 d，R2中厌氧氨氧化生物膜达到相同比厌氧氨氧化活性和血红素c含量的时间分别需要7和11 d，R3中厌氧氨氧化生物膜达到相同比厌氧氨氧化活性和血红素c含量的时间分别需要11和17 d。说明经活性恢复后的厌氧氨氧化生物膜均具有较好的脱氮效果，其中保存于4℃条件的厌氧氨氧化生物膜具有最短的微生物活性恢复时间，较为适宜保存厌氧氨氧化生物膜。
表2 保存和活性恢复后厌氧氨氧化生物膜的性状
Figure dest_path_image001
实施例4 活性恢复后厌氧氨氧化生物膜的稳定性
胞外聚合物是厌氧氨氧化生物膜形成的重要因素，而胞外聚合物中多糖类（PS）物质和蛋白质类（PN）物质的比值（PS/PN）是衡量厌氧氨氧化生物膜结构稳定的重要参数。在厌氧氨氧化生物膜活性恢复过程中，其胞外聚合物PS/PN的变化如图1所示。不同保存温度下，PS/PN差别较大，R1中厌氧氨氧化生物膜PS/PN呈先降低再升高的趋势，表明保存于-20℃条件的厌氧氨氧化生物膜稳定性逐步恢复；R3中厌氧氨氧化生物膜PS/PN经过剧烈升高再逐渐降低最后趋于稳定的过程，表明20℃不适于保存厌氧氨氧化生物膜；R2中厌氧氨氧化生物膜PS/PN总体波动较小，表明保存于4℃条件的厌氧氨氧化生物膜在活性恢复后保持较高的稳定性，4℃适宜于作为厌氧氨氧化生物膜的保存温度。
实施例5 活性恢复后厌氧氨氧化生物膜对污染物的去除效能
经过活性恢复过程后，不同保存温度下的厌氧氨氧化生物膜对氨氮和总氮的去除率均逐渐升高（图2和图3），其对氨氮和总氮的去除率分别超过90%和85%。在活性恢复的第12 d，R2中的厌氧氨氧化生物膜对氨氮和总氮的去除效果最好，并一直呈现氨氮和总氮去除率稳定升高的趋势，此结果也同表2中R2内厌氧氨氧化生物膜最快恢复较高的比厌氧氨氧化活性和血红素c含量相对应，说明4℃的条件较为适宜保存厌氧氨氧化生物膜，在实际应用中具有很高的可行性。
实施例6 活性恢复后厌氧氨氧化生物膜特性与污泥细胞状态相关性
在厌氧氨氧化生物膜活性恢复30 d后，采用流式细胞术对厌氧氨氧化生物膜细胞状态进行分析，结果如表3所示。不同保存温度下的厌氧氨氧化生物膜中活细胞含量与污水处理厂厌氧氨氧化生物膜中活细胞含量基本一致，说明经过活性恢复后，厌氧氨氧化生物膜均可发挥稳定的含氮污染物去除效果。其中R2内厌氧氨氧化生物膜活细胞比例（85.1%±5.0%）最高，且凋亡晚期细胞比例（6.1%±1.8%）和死细胞比例（3.7%±0.3%）最低，说明4℃的保存条件下厌氧氨氧化生物膜细胞活性最高，较为适宜作为保存厌氧氨氧化生物膜的条件。
表3活性恢复后厌氧氨氧化生物膜细胞活性状态
厌氧氨氧化生物膜 活细胞 凋亡早期细胞 凋亡晚期细胞 死细胞
污泥脱水滤液处理池 87.5±5.1 4.9±1.1 5.1±1.2 2.5±0.5
保存于-20℃ 83.0±4.5 5.8±1.7 6.1±1.9 5.1±1.0
保存于4℃ 85.1±5.0 5.1±1.6 6.1±1.8 3.7±0.3
保存于20℃ 80.3±5.5 5.5±1.5 8.9±1.2 5.3±1.0
如表4所示，依据Correl相关性分析发现，厌氧氨氧化生物膜比厌氧氨氧化活性和血红素c含量和厌氧氨氧化生物膜活细胞比例具有极高的相关性，相关系数分别高达0.9974和0.9930，表明利用厌氧氨氧化生物膜活细胞比例作为评价厌氧氨氧化生物膜活性的方法具有极高的可行性。同时，由于保存的厌氧氨氧化生物膜中，4℃的保存条件下厌氧氨氧化生物膜活细胞比例最高，与活性恢复后R2中厌氧氨氧化生物膜活细胞比例结果相吻合。
表4活性恢复后厌氧氨氧化生物膜特性与细胞活性状态关联性
Figure dest_path_image002
因此，确定4℃是保存厌氧氨氧化生物膜的最适温度，利用流式细胞术可以作为确定厌氧氨氧化生物膜最适保存温度的依据。流式细胞术操作简便，数据快速易得且准确可靠，亦可省略厌氧氨氧化生物膜活性恢复过程，对于厌氧氨氧化生物膜的保存与活性恢复具有重要意义。
实施例7
厌氧氨氧化生物膜保存培养：
厌氧氨氧化生物膜的保存温度设置为-20℃，4℃和20℃。将污水处理厂污泥脱水滤液处理池中的厌氧氨氧化生物膜悬浮填料150个取出，平均三等份分别置于装有750 ml保存基质的1000 ml血清瓶中（血清瓶预先充N 2以排出空气中的O 2），保存基质以NH 4Cl和NaNO 2提供NH 4 +-N和NO 2 --N，其余成分如下所示： KHCO 3 1500 mg/L, K 2HPO 4 15 mg/L，MgSO 4 180 mg/L，CaCl 2 20 mg/L；保存基质NH 4 +-N浓度为50 mg/L，NO 2 --N浓度为75 mg/L。将血清瓶（每个保存温度下设置3个平行样）分别置于-20℃，4℃和20℃，静止遮光保存。
保存的厌氧氨氧化生物膜细胞状态测试：
于-20℃，4℃和20℃保存的厌氧氨氧化生物膜在保存超过120 d后，用于测定厌氧氨氧化生物膜细胞状态，具体的流式细胞术细胞状态测试步骤如下：
（1）取10 ml厌氧氨氧化生物膜，用pH为7.8的磷酸盐缓冲液稀释至100 ml，于涡旋仪漩涡震荡2 min，使生物膜破碎为絮体并保证均匀分布；
（2）用8μm孔径的尼龙膜过滤破碎后的样品，取1.5 ml置于1.5 ml尖底离心管中；
（3）样品于8000 rpm/min 离心5 min；
（4）用移液枪吸取离心后的样品上清液，留下约0.1 ml样品，使用预冷磷酸盐缓冲液（pH为7.8）吹洗细胞，重复离心和清洗两次；
（5）离心后的样品用移液枪吸取上清液，留下约0.1 ml样品，使用0.3 ml 10x Annexin V Binding Buffer混匀；
（6）对照FITC Annexin V组加入0.5μl PI染色剂，对照PI组加入0.5 μl FITC Annexin V，检测组加入0.5μl FITC Annexin V和0.5 μl PI，混匀后室温下避光孵育15 min，之后于流式细胞仪上机检测。
厌氧氨氧化生物膜细胞状态测试结果如表5所示。
表5 保存120 d的厌氧氨氧化生物膜细胞活性状态
Figure dest_path_image003
由表5结果发现，选用“pH为7.8的磷酸盐缓冲液获得”所测得的细胞活性差异度明显弱于选用“pH为7.8的磷酸盐缓冲液和胎牛血清混合后的缓冲液”的活性数据结果，且选用pH为7.8的磷酸盐缓冲液时，各活性数据的数值偏差较大、波动范围过宽，会显著影响流式细胞术结果。结合不同保存温度下细胞活性数据较弱的差异性，最终易引起较大的偏差，造成测试结果与验证结果不吻合，无法有效反应通过流式细胞术作为保存温度判断的依据。
同时，发明人还发现缓冲液pH对测试过程中的细胞状态影响较大，具有较好的细胞活性数据分析效力的是弱碱性pH7.2~8.0的环境，中性偏酸或者碱性过大条件下的细胞状态数据差距较小，无法准确判断结果。
此外，发明人还分别考察了6μm和10μm孔径制得的样品，6μm样品测试结果与验证实验一致，数据可靠；而10μm相应数据不具备分析效力，无法用来确定最适合保存温度。
虽然本发明已以较佳实施例公开如上，但其并非用以限定本发明，任何熟悉此技术的人，在不脱离本发明的精神和范围内，都可做各种的改动与修饰，因此本发明的保护范围应该以权利要求书所界定的为准。

Claims (12)

  1. 一种确认污水处理厌氧氨氧化生物膜最适保存温度的方法，其特征在于，所述方法是基于流式细胞术对厌氧氨氧化生物膜的细胞活性状态进行测定，对比不同温度保存的厌氧氨氧化生物膜与保存前厌氧氨氧化生物膜的细胞活性状态的测定结果，以最接近保存前厌氧氨氧化生物膜的细胞活性状态时保存的温度作为最适保存温度；所述厌氧氨氧化生物膜的细胞活性状态测定包括活细胞、凋亡早期细胞、凋亡晚期细胞和死细胞的含量测定；
    所述流式细胞术确认最适温度的步骤包括：
    （1）厌氧氨氧化生物膜测试样液的制备：用缓冲液稀释厌氧氨氧化生物样品，震荡均匀后，过滤、离心，留用上清液，使用预冷磷酸盐缓冲液吹洗细胞，重复离心和清洗两次，再取上清液作为样品，使用适量10x Annexin V Binding Buffer混匀制得；其中，缓冲液包括磷酸盐缓冲液和胎牛血清，缓冲液的pH为7.2~8.0，与厌氧氨氧化生物膜的稀释比为8~10：1；
    （2）置于流式细胞仪测定各样液的细胞活性状态。
  2. 根据权利要求1所述方法，其特征在于，所述过滤是选用6~8 μm孔径的尼龙膜。
  3. 一种确认污水处理厌氧氨氧化生物膜最适保存温度的方法，其特征在于，所述方法是基于流式细胞术对厌氧氨氧化生物膜的细胞活性状态进行测定，对比不同温度保存的厌氧氨氧化生物膜与保存前厌氧氨氧化生物膜的细胞活性状态的测定结果，以最接近保存前厌氧氨氧化生物膜的细胞活性状态时保存的温度作为最适保存温度；所述厌氧氨氧化生物膜的细胞活性状态测定包括活细胞、凋亡早期细胞、凋亡晚期细胞和死细胞的含量测定。
  4. 根据权利要求3所述方法，其特征在于，所述流式细胞术确认最适温度的步骤包括：
    （1）厌氧氨氧化生物膜测试样液的制备：用缓冲液稀释厌氧氨氧化生物样品，震荡均匀后，过滤、离心，留用上清液，使用预冷磷酸盐缓冲液吹洗细胞，重复离心和清洗两次，再取上清液作为样品，使用适量10x Annexin V Binding Buffer混匀制得；
    （2）置于流式细胞仪测定各样液的细胞活性状态。
  5. 根据权利要求4所述方法，其特征在于，所述缓冲液包括磷酸盐缓冲液和胎牛血清。
  6. 根据权利要求4或5所述方法，其特征在于，所述磷酸盐缓冲液包括0.2 mol/L磷酸氢二钠和0.2 mol/L磷酸二氢钠。
  7. 根据权利要求4或5所述方法，其特征在于，所述缓冲液中的磷酸盐缓冲液和胎牛血清的体积比为8~10：1。
  8. 根据权利要求4或5所述方法，其特征在于，所述缓冲液的pH为7.2~8.0，与厌氧氨氧化生物膜的稀释比为8~10：1。
  9. 根据权利要求4或5任一所述方法，其特征在于，所述过滤是选用6~8 μm孔径的尼龙膜。
  10. 一种快速启动厌氧氨氧化生物膜污水处理的方法，其特征在于，利用权利要求1所述方法测得最适保存温度，具体包括：预先将厌氧氨氧化生物膜培养成熟，置于保存基质中在最适保存温度下进行保存，活性恢复后即可用于污水处理。
  11. 根据权利要求10所述方法，其特征在于，所述保存基质包括：KHCO 3 1500 mg/L, K 2HPO 4 15 mg/L，MgSO 4 180 mg/L，CaCl 2 20 mg/L；保存基质NH 4 +-N浓度为50 mg/L，NO 2 --N浓度为75 mg/L。
  12. 权利要求3或10所述方法在污水处理中的应用。
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2009058233A (ja) * 2007-08-29 2009-03-19 Nagasaki Prefecture 入浴設備の汚染度の判定方法、浴槽水における殺微生物剤の殺微生物効果の判定方法及び浴槽水の水質管理方法
CN103293138A (zh) * 2013-06-21 2013-09-11 北京大学 一种快速表征剩余污泥碱性厌氧发酵过程与产物的方法
CN103808703A (zh) * 2014-02-27 2014-05-21 湖南大学 测定处理废水后黄孢原毛平革菌菌体内活性氧水平的方法
CN104845997A (zh) * 2015-02-13 2015-08-19 温州医科大学 一株检测铜的大肠埃希氏菌
CN106635803A (zh) * 2017-01-06 2017-05-10 大连理工大学 一种厌氧氨氧化菌干粉菌剂制备与保藏的方法

Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5651891A (en) * 1989-08-02 1997-07-29 Polytechnic University Wastewater treatment process
US6998234B2 (en) * 2000-11-03 2006-02-14 Oncotech, Inc. Methods for cancer prognosis and diagnosis relating to tumor vascular endothelial cells
JP4284700B2 (ja) * 2004-03-25 2009-06-24 株式会社日立プラントテクノロジー 窒素除去方法及び装置
US7314563B2 (en) * 2005-11-14 2008-01-01 Korea Institute Of Science And Technology Membrane coupled activated sludge method and apparatus operating anoxic/anaerobic process alternately for removal of nitrogen and phosphorous
BRPI0721832B1 (pt) * 2007-08-08 2018-03-20 Peng Guanghao "método para a remoção de contaminantes de carbono e nitrogênio de efluentes usando bactérias heterotróficas oxidantes da amônia"
JP5459209B2 (ja) * 2008-07-11 2014-04-02 星光Ｐｍｃ株式会社 微生物群集の生理状態の判定方法及び排水処理方法
CN101418295B (zh) * 2008-12-11 2010-12-22 中国科学院微生物研究所 一种生物传感器及其应用
US8864993B2 (en) * 2012-04-04 2014-10-21 Veolia Water Solutions & Technologies Support Process for removing ammonium from a wastewater stream
CN103755109B (zh) * 2013-12-29 2015-04-22 杭州师范大学 一种厌氧氨氧化颗粒污泥的保存和重激活方法
CN111003813B (zh) * 2018-10-29 2021-05-28 江南大学 一种确定硫自养反硝化菌生物膜最适保存温度的方法

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2009058233A (ja) * 2007-08-29 2009-03-19 Nagasaki Prefecture 入浴設備の汚染度の判定方法、浴槽水における殺微生物剤の殺微生物効果の判定方法及び浴槽水の水質管理方法
CN103293138A (zh) * 2013-06-21 2013-09-11 北京大学 一种快速表征剩余污泥碱性厌氧发酵过程与产物的方法
CN103808703A (zh) * 2014-02-27 2014-05-21 湖南大学 测定处理废水后黄孢原毛平革菌菌体内活性氧水平的方法
CN104845997A (zh) * 2015-02-13 2015-08-19 温州医科大学 一株检测铜的大肠埃希氏菌
CN106635803A (zh) * 2017-01-06 2017-05-10 大连理工大学 一种厌氧氨氧化菌干粉菌剂制备与保藏的方法

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