CN114890537A - Filamentous sludge bulking control method based on quorum sensing - Google Patents
Filamentous sludge bulking control method based on quorum sensing Download PDFInfo
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- 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
- C02F3/12—Activated sludge processes
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- 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
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
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Abstract
The invention discloses a filamentous sludge bulking control method based on quorum sensing. Belongs to the technical field of filamentous sludge bulking control. The method comprises the steps of operating an activated sludge reactor, collecting an activated sludge sample, judging the sludge bulking degree and the sludge bulking type, performing gram staining and Neisseria staining, identifying the dominant filamentous bacteria causing filamentous sludge bulking, determining the types and the contents of N-Acylated Homoserine Lactone (AHLs) signal molecules, performing correlation analysis to determine the AHLs signal molecules which are negatively correlated with the filamentous sludge bulking, and adding the determined AHLs signal molecules into the activated sludge reactor, thereby realizing the control of the filamentous sludge bulking in the activated sludge reactor. The method is simple and easy to operate, can quickly reduce or eliminate the negative effects of difficult sludge sedimentation, deteriorated effluent quality and the like caused by sludge bulking, and improves the operation stability and the operation safety of the conventional sewage treatment plant.
Description
Technical Field
The invention relates to the technical field of filamentous sludge bulking control, in particular to a filamentous sludge bulking control method based on quorum sensing.
Background
The activated sludge process has become the mainstream technology in the field of sewage treatment at present. However, as a persistent problem in the activated sludge process, the sludge bulking phenomenon has been an important problem affecting the stable operation of the sewage treatment system. In the study on the sludge bulking phenomenon, it was found that more than 90% of the sludge bulking problem was caused by the overgrowth of filamentous fungi. Therefore, the control of the filamentous sludge bulking plays an important role in the normal operation of sewage treatment plants.
The existing commonly used filamentous sludge bulking control method is to add chemical agents such as oxidant and the like to control the growth of filamentous bacteria, but the method kills the filamentous bacteria and damages other functional bacteria in the system at the same time, so that the ecological system in the system is unbalanced; in addition, the sludge bulking can be controlled by regulating and controlling the process running conditions such as sludge load, sludge age, sludge reflux ratio and the like, but the methods have long period and slow effect and are not beneficial to practical application.
The activated sludge is taken as an ecological system with multi-flora aggregation, which mainly depends on a large amount of microorganisms to remove pollutants so as to realize the stable operation of a sewage treatment system, and the filamentous sludge bulking is mainly caused by the disruption of flora ecological balance caused by the overgrowth of filamentous bacteria. Quorum sensing is a phenomenon widely existing in various microorganisms, and is a phenomenon that cell-cell communication is realized by generating sensing chemical signal molecules, so that the behavior of the quorum is controlled. When the signal molecules reach the threshold concentration, the homologous receptors combine the signal molecules and trigger signal transduction cascade to stimulate the expression of related genes in the bacteria, regulate the physiological behavior of the flora and the ecological relationship of the flora, and finally determine the population structure, thereby showing the physiological function and the regulation mechanism which can not be realized by single individual bacteria. However, there are few reports on methods for controlling filamentous sludge bulking based on quorum sensing in the prior art.
Therefore, from the quorum sensing angle of the filamentous fungi in the activated sludge system, an effective control method of filamentous sludge bulking based on quorum sensing regulation is developed, and the method has important significance for sludge bulking of a sewage treatment plant.
Disclosure of Invention
In view of this, the invention provides a filamentous sludge bulking control method based on quorum sensing. The method analyzes the species and the content of filamentous bacteria and N-Acylated Homoserine Lactones (AHLs) signal molecules which initiate sludge bulking by analyzing a sample which generates sludge bulking, determines the AHLs signal molecules which can inhibit sludge bulking and then adds the signal molecules externally, and realizes the control of filamentous sludge bulking.
In order to achieve the purpose, the invention adopts the following technical scheme:
a filamentous sludge bulking control method based on quorum sensing comprises the following steps:
(1) operating the activated sludge reactor;
(2) collecting an activated sludge sample in the activated sludge reactor in the step (1), measuring a Sludge Volume Index (SVI) of the activated sludge sample, and carrying out microscope in-situ observation to determine the sludge bulking degree and the sludge bulking type of the activated sludge sample;
(3) performing gram staining and Neisseria staining on the activated sludge sample obtained in the step (2), and identifying dominant filamentous bacteria which cause filamentous sludge bulking;
(4) measuring the species and the content of N-Acylated Homoserine Lactones (AHLs) signal molecules of the activated sludge sample obtained in the step (2);
(5) performing correlation analysis on the AHLs signal molecules detected in the step (4) and filamentous sludge bulking to determine the AHLs signal molecules which are negatively correlated with the filamentous sludge bulking;
(6) and (4) adding the AHLs signal molecules which are determined in the step (5) and are negatively related to filamentous sludge bulking into the activated sludge reactor, so as to realize the control of the filamentous sludge bulking in the activated sludge reactor.
The mechanism is as follows: the filamentous fungi senses the concentration of extracellular AHLs signal molecules in the system, when the concentration of the extracellular AHLs signal molecules is high, corresponding receptor proteins in the cells of the filamentous fungi receive signals to regulate and control corresponding genes not to be expressed, so that the physiological behavior of the filamentous fungi is regulated and controlled, the growth of the filamentous fungi is inhibited, and the aim of controlling the expansion of filamentous sludge is fulfilled.
The advantages are that: the existing commonly used filamentous sludge bulking control method is to add chemical agents such as oxidant and the like to control the growth of filamentous bacteria, but the method kills the filamentous bacteria and damages other functional bacteria in the system at the same time, so that the ecological system in the system is unbalanced; in addition, the sludge bulking can be controlled by regulating and controlling the process running conditions such as sludge load, sludge age, sludge reflux ratio and the like, but the methods have long period and slow effect and are not beneficial to practical application. The method of the invention makes full use of the quorum sensing regulation mechanism among microorganisms, and according to the correlation relationship between AHLs signal molecules and filamentous sludge bulking, the AHLs signal molecules for inhibiting the growth of filamentous bacteria are added in a targeted manner, thereby realizing the control of the sludge bulking. The method is simple to operate and convenient to implement, and can quickly realize filamentous sludge bulking control and stable system operation.
Further, the dissolved oxygen concentration of the activated sludge reactor in the step (1) is controlled to be 1.5-2.0 mg/L during the aerobic aeration.
Further, the hydraulic retention time of the activated sludge reactor in the step (1) is 8-20 h.
Further, in the step (1), the activated sludge reactor is filled with water, artificial preparation of simulated wastewater is adopted, sodium acetate is used as a carbon source, ammonium sulfate is used as a nitrogen source, dipotassium hydrogen phosphate is used as a phosphorus source, the COD concentration of the artificial preparation of the simulated wastewater is 250-350 Mg/L, the ammonia nitrogen concentration is 50-70 Mg/L, and the phosphorus concentration is 5-7 Mg/L, magnesium sulfate, calcium chloride, ferric chloride, boric acid, cobalt chloride, nickel chloride, zinc sulfate, manganese chloride and copper sulfate are added to provide constant and trace elements, so that the concentrations of the constant and trace elements are respectively Mg 2+ :2mg/L,Ca 2+ :10mg/L,Fe 3+ :0.3mg/L,B 3+ :30μg/L,Co 2+ :35μg/L,Ni + :35μg/L,Zn 2+ :30μg/L,Mn 2 + :30μg/L,Cu 2+ : mu.g/L, and sodium bicarbonate was added to adjust the pH to 8.0.
The beneficial effects are as follows: the setting not only meets the requirement of normal growth of the microorganism, but also does not have the stress effect on the microorganism in the system due to overhigh concentration of COD, nitrogen and phosphorus. The above concentration settings refer to the domestic sewage concentration in a sewage treatment plant.
Further, the concentration of sludge in the activated sludge reactor in the step (1) is controlled to be 3000-3500 mg/L, and the sludge age is controlled to be 20-25 days.
The beneficial effects are as follows: the sludge concentration is too high, so that the sludge load is reduced, the sludge concentration is too low, the sludge load is high, and the sludge load is too low or too high, which is unfavorable for the system operation; if the sludge age is too short, the generation time of partial functional bacteria such as ammonia oxidizing bacteria and nitrite oxidizing bacteria is long, and the growth of the partial functional bacteria is not facilitated, and if the sludge age is too long, the phosphorus-accumulating sludge is discharged less, and phosphorus-accumulating bacteria is not facilitated to remove phosphorus.
Further, in the step (1), the temperature of the activated sludge reactor is controlled to be 20-25 ℃.
The beneficial effects are as follows: various microorganisms can normally grow and keep higher activity in the temperature range, sludge expansion is easy to cause if the temperature is too low, and meanwhile, partial functional bacteria are not beneficial to growth, and partial enzyme activity of partial microorganisms is inhibited if the temperature is too high, so that the normal growth of the microorganisms is not beneficial.
Furthermore, the SVI determination in the step (2) should be performed immediately after sampling, and the microscope in-situ observation should be performed within 0-2 h.
Further, the AHLs signal molecules in the step (4) are extracted by a solid phase extraction method and are measured by a high performance liquid chromatography-mass spectrometer.
Further, the concentration of the AHLs signal molecules which are negatively related to the filamentous sludge bulking is 5 ng/L-25 ng/L after the AHLs signal molecules are added in the step (6).
The beneficial effects are as follows: the concentration is effective concentration for regulating and controlling the expansion of filamentous sludge.
Further, when the SVI value of the activated sludge reactor in the step (1) is stabilized below 150mL/g, the AHLs signal molecules which are negatively related to the filamentous sludge bulking in the step (6) are not added.
According to the technical scheme, compared with the prior art, the invention has the following beneficial effects: the operation is simple and easy, the negative effects of difficult sludge sedimentation, deteriorated effluent quality and the like caused by sludge bulking can be quickly reduced or eliminated, and the operation stability and the operation safety of the existing sewage treatment plant are improved. The invention controls sludge bulking from the perspective of quorum sensing regulation, effectively and reasonably utilizes the ecological effects of filamentous bacteria and other microbial communities, and can quickly realize the control of filamentous sludge bulking and the stable operation of a system.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 is a gram stain plot of a sludge sample when filamentous sludge bulking occurs in example 1 of the present invention;
FIG. 2 is a graph showing the change of SVI and AHLs signal molecules during the operation of SBR reactor in example 1 of the present invention;
FIG. 3 is a graph showing the change in SVI after C6-HSL is added in example 1 of the present invention;
FIG. 4 is a gram stain chart of a sludge sample after filamentous sludge bulking control in example 1 of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The required medicament is a conventional experimental medicament purchased from a market channel; the unrecited experimental method is a conventional experimental method, and is not described in detail herein.
Example 1
The activated sludge reactor operated in the example is an SBR reactor, the effective volume is 4L, after the secondary sedimentation tank is inoculated with the return sludge, the sludge concentration is 3200mg/L, and the SVI is 94 mL/g. Operating in an anoxic/aerobic manner, and operating for 4 periods every day, wherein water is fed into each period for 10min, anaerobic/anoxic stirring is performed for 110min, aerobic aeration is performed for 180min, precipitation is performed for 50min, water is discharged for 7min, and the operation is idle for 3 min. The SBR reactor is controlled by a constant-temperature water area interlayer to have the temperature of 20 +/-1 ℃, the dissolved oxygen concentration is controlled to be 1.0-1.5 mg/L during aerobic aeration, water inlet and water discharge are controlled by a peristaltic pump, the water discharge ratio is 50%, the hydraulic retention time is 12h, and the sludge age is 25 days. During the operation period, the water inlet of the SBR reactor adopts artificial preparation of simulated wastewater, sodium acetate is used as a carbon source, ammonium sulfate is used as a nitrogen source, dipotassium hydrogen phosphate is used as a phosphorus source, the COD concentration is 300mg/L, the ammonia nitrogen concentration is 60mg/L, the phosphorus concentration is 5mg/L, and sodium bicarbonate is added to adjust the pH value to 8.0. In addition, magnesium sulfate, calcium chloride, ferric chloride, boric acid, cobalt chloride, nickel chloride, zinc sulfate, manganese chloride and copper sulfate are added to provide major and trace elements with concentrations of Mg 2+ :2mg/L,Ca 2+ :10mg/L,Fe 3+ :0.3mg/L,B 3+ :30μg/L,Co 2+ :35μg/L,Ni + :35μg/L,Zn 2+ :30μg/L,Mn 2+ :30μg/L,Cu 2+ :8μg/L。
Taking an activated sludge sample in the SBR reactor for SVI (singular value analysis), wherein the SVI comprises the following steps: taking 100mL of activated sludge mixed liquor, standing for 30min, and recording SV of the mixed liquor 30 Then filtering with qualitative filter paper, putting into a 105 ℃ oven to dry to constant weight to obtain the dry weight W of the sludge, wherein the dry weight is SV 30 and/W to obtain SVI. After 12 days of operation, the SVI increased from 94mL/g to 152mL/g, and the presence of filamentous bacteria was observed in situ with a microscope, at which time the system had swollen filamentous sludge. Then, taking an activated sludge sample for gram staining, wherein the staining method comprises the following steps: diluting 5mL of activated sludge mixed liquor by 3 times, uniformly coating 100 mu L of diluted mixed liquor on a glass slide, after the glass slide is air-dried, primarily dyeing the glass slide for 1min by using an ammonium oxalate crystal violet solution, mordanting iodine solution for 1min, decoloring the glass slide by using 95% ethanol for 30s, and re-dyeing safranin dyeing liquor for 1 min. Observing the gram-stained sample by using a microscope, wherein filamentous bacteria are gram-positive, have slender and smooth hyphae and are in line with morphological characteristics of the microfilaria, and the dominant filamentous bacteria causing filamentous sludge bulking are identified to be the microfilaria according to the morphological characteristics (as shown in figure 1). Meanwhile, a solid phase extraction method is adopted to extract AHLs signal molecules in an activated sludge sample, and the steps of the solid phase extraction are as follows: respectively activating a Bond Elut ENV solid phase extraction column by using 5mL of chromatographic methanol and 5mL of pure water, loading 200mL of a water sample filtered by a 0.45-micron filter membrane into the solid phase extraction column at a speed of 5-10 mL/min, then eluting by using 5mL of 5% methanol aqueous solution, eluting an extracted AHLs signal molecule by using 10mL of chromatographic methanol after the solid phase extraction column is drained, collecting an eluent, desolventizing the eluent by using a rotary evaporator, and fixing the volume to 2mL by using an initial mobile phase. And determining the types and the contents of AHLs signal molecules by using a high performance liquid chromatography-mass spectrometer, wherein the specific determination conditions are as follows. High performance liquid chromatography conditions: the flow rate is 0.2mL/min, the sample amount is 10 mul, the mobile phase A is aqueous solution containing 2mM ammonium acetate and 0.1% formic acid, the mobile phase B is methanol, the total analysis time is 10min, the operation is carried out for 1min, and the gradient elution condition is adopted to obtain good separation effect. Mass spectrometryConditions are as follows: adopting a positive ion scanning mode, wherein the ion source is an electrospray ionization source (AJS-ESI), and the monitoring mode is a multi-reaction monitoring (MRM) mode; the temperature of the drying gas is 350 ℃, and the flow rate is 8L/min; the atomizer pressure was 30psi, the sheath gas temperature was 350 deg.C, the sheath gas flow rate was 11L/min, and the capillary voltage was 3500V. Thereafter, samples were taken every 2 days to determine SVI and AHLs signal molecules (see FIG. 2), and gram-stained microscopic observations were made, and Pearson correlation analysis was performed using SPSS as variables for SVI and AHLs signal molecule concentration, respectively, to obtain C6-HSL which exhibited a significant negative correlation (p < 0.01) with SVI, and a Pearson correlation coefficient of-0.93.
After the SBR reactor is operated for 40 days, C6-HSL is added into the system, the mixture is added into the SBR reactor with the inflow water, and the concentration of the C6-HSL is 15ng/L after the mixture is added. After 2 days of dosing (i.e., 42 days of system operation), the SVI decreased from 284mL/g to 273mL/g, and by day 64, the SVI decreased to 147mL/g, and thereafter stabilized below 150mL/g (FIG. 3), at which time the gram stain plot (FIG. 4) showed that the sludge flocs were structurally compact, the microfilaments were few and short and present in the sludge flocs, and empty cells were observed in the middle of the hyphae, indicating that the microfilaments were about to die. And stopping adding the signal molecule C6-HSL on the 70 th day, and continuing running for 20 days, wherein the SVI is stabilized below 150 mL/g.
The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (10)
1. A filamentous sludge bulking control method based on quorum sensing is characterized by comprising the following steps:
(1) operating the activated sludge reactor;
(2) collecting an activated sludge sample in the activated sludge reactor in the step (1), measuring the SVI of the activated sludge sample, and carrying out in-situ observation by using a microscope to determine the sludge bulking degree and the sludge bulking type of the activated sludge sample;
(3) performing gram staining and Neisseria staining on the activated sludge sample obtained in the step (2), and identifying dominant filamentous bacteria which cause filamentous sludge bulking;
(4) measuring the species and content of AHLs signal molecules of the activated sludge sample obtained in the step (2);
(5) performing correlation analysis on the AHLs signal molecules detected in the step (4) and filamentous sludge bulking to determine the AHLs signal molecules which are negatively correlated with the filamentous sludge bulking;
(6) and (4) adding the AHLs signal molecules which are determined in the step (5) and are negatively related to filamentous sludge bulking into the activated sludge reactor, so as to realize the control of the filamentous sludge bulking in the activated sludge reactor.
2. The method for controlling filamentous sludge bulking based on quorum sensing as claimed in claim 1, wherein the dissolved oxygen concentration of the activated sludge reactor in step (1) during aerobic aeration is controlled to be 1.5-2.0 mg/L.
3. The filamentous sludge bulking control method based on quorum sensing as claimed in claim 1, wherein the hydraulic retention time of the activated sludge reactor in step (1) is 8-20 h.
4. The filamentous sludge bulking control method based on quorum sensing as claimed in claim 1, wherein in step (1), the activated sludge reactor is fed with water to artificially prepare simulated wastewater, sodium acetate is used as a carbon source, ammonium sulfate is used as a nitrogen source, dipotassium hydrogen phosphate is used as a phosphorus source, the COD concentration of the artificially prepared simulated wastewater is 250-350 mg/L, the ammonia nitrogen concentration is 50-70 mg/L, and the phosphorus concentration is 5-7 mg/L, and meanwhile, the artificially prepared simulated wastewater has the COD concentration of 250-350 mg/L, the ammonia nitrogen concentration is 50-70 mg/L, and the phosphorus concentration is 5-7 mg/LMagnesium sulfate, calcium chloride, ferric chloride, boric acid, cobalt chloride, nickel chloride, zinc sulfate, manganese chloride and copper sulfate are added to provide major elements and trace elements, so that the concentrations of the major elements and the trace elements are respectively Mg 2+ :2mg/L,Ca 2+ :10mg/L,Fe 3 + :0.3mg/L,B 3+ :30μg/L,Co 2+ :35μg/L,Ni + :35μg/L,Zn 2+ :30μg/L,Mn 2+ :30μg/L,Cu 2+ : mu.g/L, and sodium bicarbonate was added to adjust the pH to 8.0.
5. The filamentous sludge bulking control method based on quorum sensing as claimed in claim 1, wherein in step (1), the activated sludge reactor is controlled to have a sludge concentration of 3000-3500 mg/L and a sludge age of 20-25 days.
6. The quorum sensing-based filamentous sludge bulking control method according to claim 1, wherein the temperature of the activated sludge reactor in the step (1) is controlled to be 20-25 ℃.
7. The method for controlling filamentous sludge bulking based on quorum sensing as claimed in claim 1, wherein the SVI determination in step (2) is performed immediately after sampling, and the microscopic in situ observation is performed within 0-2 h.
8. The filamentous sludge bulking control method based on quorum sensing of claim 1, wherein in step (4) the AHLs signal molecules are extracted by solid phase extraction and measured by high performance liquid chromatography-mass spectrometry.
9. The filamentous sludge bulking control method based on quorum sensing as claimed in claim 1, wherein the concentration of AHLs signal molecules negatively correlated to filamentous sludge bulking in step (6) is 5 ng/L-25 ng/L.
10. The method for controlling filamentous sludge bulking based on quorum sensing as claimed in claim 1, wherein when the SVI value of the activated sludge reactor in step (1) is stabilized below 150mL/g, the AHLs signal molecules negatively correlated to filamentous sludge bulking in step (6) are not added.
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CN110117078A (en) * | 2019-04-03 | 2019-08-13 | 东北师范大学 | A kind of filamentous fungi property sludge bulking inhibitor and its application method |
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CN110117078A (en) * | 2019-04-03 | 2019-08-13 | 东北师范大学 | A kind of filamentous fungi property sludge bulking inhibitor and its application method |
CN112978907A (en) * | 2021-02-09 | 2021-06-18 | 北京工业大学 | Device and method for quickly starting filamentous bacterium sludge expansion under low-temperature condition |
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