CN114426335B - Co-enrichment method of nitrifying bacteria and denitrifying phosphorus accumulating bacteria - Google Patents
Co-enrichment method of nitrifying bacteria and denitrifying phosphorus accumulating bacteria Download PDFInfo
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- 241000894006 Bacteria Species 0.000 title claims abstract description 116
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 title claims abstract description 85
- 229910052698 phosphorus Inorganic materials 0.000 title claims abstract description 85
- 239000011574 phosphorus Substances 0.000 title claims abstract description 85
- 238000000034 method Methods 0.000 title claims abstract description 72
- 230000001546 nitrifying effect Effects 0.000 title claims abstract description 56
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 22
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- 239000010802 sludge Substances 0.000 claims description 30
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 claims description 26
- 239000000758 substrate Substances 0.000 claims description 25
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 21
- 229910052760 oxygen Inorganic materials 0.000 claims description 21
- 239000001301 oxygen Substances 0.000 claims description 21
- 238000003756 stirring Methods 0.000 claims description 21
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- 238000004062 sedimentation Methods 0.000 claims description 7
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- KWIUHFFTVRNATP-UHFFFAOYSA-O N,N,N-trimethylglycinium Chemical compound C[N+](C)(C)CC(O)=O KWIUHFFTVRNATP-UHFFFAOYSA-O 0.000 claims description 6
- 229960003237 betaine Drugs 0.000 claims description 6
- WGTYBPLFGIVFAS-UHFFFAOYSA-M tetramethylammonium hydroxide Chemical compound [OH-].C[N+](C)(C)C WGTYBPLFGIVFAS-UHFFFAOYSA-M 0.000 claims description 6
- 238000007599 discharging Methods 0.000 claims description 5
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 4
- 239000004475 Arginine Substances 0.000 claims description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 4
- PMMYEEVYMWASQN-DMTCNVIQSA-N Hydroxyproline Chemical compound O[C@H]1CN[C@H](C(O)=O)C1 PMMYEEVYMWASQN-DMTCNVIQSA-N 0.000 claims description 4
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 4
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 4
- PMMYEEVYMWASQN-UHFFFAOYSA-N dl-hydroxyproline Natural products OC1C[NH2+]C(C([O-])=O)C1 PMMYEEVYMWASQN-UHFFFAOYSA-N 0.000 claims description 4
- 229960002591 hydroxyproline Drugs 0.000 claims description 4
- FGMPLJWBKKVCDB-UHFFFAOYSA-N trans-L-hydroxy-proline Natural products ON1CCCC1C(O)=O FGMPLJWBKKVCDB-UHFFFAOYSA-N 0.000 claims description 4
- 229930006000 Sucrose Natural products 0.000 claims description 3
- 239000005720 sucrose Substances 0.000 claims description 3
- ZTOKUMPYMPKCFX-CZNUEWPDSA-N (E)-17-[(2R,3R,4S,5S,6R)-6-(acetyloxymethyl)-3-[(2S,3R,4S,5S,6R)-6-(acetyloxymethyl)-3,4,5-trihydroxyoxan-2-yl]oxy-4,5-dihydroxyoxan-2-yl]oxyoctadec-9-enoic acid Chemical compound OC(=O)CCCCCCC/C=C/CCCCCCC(C)O[C@@H]1O[C@H](COC(C)=O)[C@@H](O)[C@H](O)[C@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](COC(C)=O)O1 ZTOKUMPYMPKCFX-CZNUEWPDSA-N 0.000 claims description 2
- 239000004471 Glycine Substances 0.000 claims description 2
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 claims description 2
- 239000002054 inoculum Substances 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 11
- 230000002195 synergetic effect Effects 0.000 abstract description 3
- 241000699670 Mus sp. Species 0.000 abstract description 2
- 238000011065 in-situ storage Methods 0.000 abstract description 2
- 244000005700 microbiome Species 0.000 description 9
- 230000000052 comparative effect Effects 0.000 description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 229910002651 NO3 Inorganic materials 0.000 description 3
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 3
- 239000003344 environmental pollutant Substances 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 231100000719 pollutant Toxicity 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 2
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 230000001651 autotrophic effect Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000001590 oxidative effect Effects 0.000 description 2
- 229920002627 poly(phosphazenes) Polymers 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 238000002798 spectrophotometry method Methods 0.000 description 2
- 230000001360 synchronised effect Effects 0.000 description 2
- 239000002351 wastewater Substances 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 108090000913 Nitrate Reductases Proteins 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- APUPEJJSWDHEBO-UHFFFAOYSA-P ammonium molybdate Chemical compound [NH4+].[NH4+].[O-][Mo]([O-])(=O)=O APUPEJJSWDHEBO-UHFFFAOYSA-P 0.000 description 1
- 229940010552 ammonium molybdate Drugs 0.000 description 1
- 235000018660 ammonium molybdate Nutrition 0.000 description 1
- 239000011609 ammonium molybdate Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000033116 oxidation-reduction process Effects 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000004886 process control Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Classifications
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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
-
- 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
- C02F3/308—Biological phosphorus removal
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
- C12N1/20—Bacteria; Culture media therefor
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- 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
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
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- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2209/00—Controlling or monitoring parameters in water treatment
- C02F2209/02—Temperature
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- 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
- C02F2209/06—Controlling or monitoring parameters in water treatment pH
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- 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
- C02F2209/08—Chemical Oxygen Demand [COD]; Biological Oxygen Demand [BOD]
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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- C02F2209/14—NH3-N
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- C02F2209/22—O2
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- 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
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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
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- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
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Abstract
The invention discloses a common enrichment method of nitrifying bacteria and denitrifying phosphorus accumulating bacteria, which comprises a nitrifying bacteria culture stage and a denitrifying phosphorus accumulating bacteria culture stage which are repeated for a plurality of times, wherein free amino acid is added in the nitrifying bacteria culture process, mice Li Tangzhi and quaternary ammonium base are added under different conditions in the denitrifying phosphorus accumulating bacteria culture process, and the culture is finished by controlling the culture conditions until the relative abundance of nitrifying bacteria and denitrifying phosphorus accumulating bacteria in the cultured flora is more than 15 percent. The nitrifying bacteria and the denitrifying phosphorus accumulating bacteria obtained by the method have high activity, good synergistic effect and strong adaptability, can be used in situ by on-site culture, and are particularly suitable for solving the problem that the total nitrogen and the total phosphorus in the existing sewage treatment field do not reach the standard.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a method for jointly enriching nitrifying bacteria and denitrifying phosphorus accumulating bacteria.
Background
Traditional biological denitrification and dephosphorization of sewage is mainly finished by nitrifying bacteria, denitrifying bacteria, phosphorus accumulating bacteria and the like. Wherein nitrifying bacteria belong to aerobic autotrophic microorganisms, and are mainly used for oxidizing ammonia into nitrite and nitrate, and under the condition of sufficient supply, energy is obtained in the process of oxidizing ammonia and nitrite so as to assimilate CO 2 The growth is slow, the generation time is long, and the growth is easily influenced by a plurality of factors such as substrate concentration, dissolved Oxygen (DO) concentration, pH and the like, so that the proliferation speed is low, the rapid culture is not easy, and the industrial large-scale application is difficult. Denitrifying bacteria are mostly heterotrophic facultative bacteria that convert nitrate to nitrogen. The phosphorus accumulating bacteria are heterotrophic microorganisms, and mainly suck phosphorus in sewage into the bacteria by virtue of that the aerobic phosphorus uptake during growth and proliferation is larger than the phosphorus release during anaerobic process, so that the residual sewage containing phosphorus can be dischargedSludge to achieve dephosphorization.
The traditional biological denitrification and dephosphorization process is difficult to realize the stability of system operation and the efficient removal of pollutants in the same reactor because of the difference between microorganisms, so that in order to achieve better removal effect, most microorganisms with different functions are placed in different reactors, so that the system has long flow, multiple structures and complex operation management. For example, CN201710146749.0 discloses a biological filter for anaerobic ammoxidation coupling denitrification dephosphorization and an operation method thereof. CN201710269314.5 discloses a sewage treatment device and method for denitrification dephosphorization tandem integrated anaerobic ammoxidation. CN201510393569.3 discloses a method for simultaneous denitrification and dephosphorization by SBR partial denitrification and dephosphorization/anaerobic ammoxidation. Each microorganism related to the method plays a role in a separate reactor, and the operation management is complex.
With the continuous and intensive research on sewage treatment, novel denitrification and dephosphorization processes and related microorganisms are developed and applied. In particular, the denitrification dephosphorization process ensures that two independent processes of dephosphorization and denitrification can be completed simultaneously only in an anoxic environment under the participation of denitrification dephosphorization bacteria (DPB), the combination of the phosphorus absorption and denitrification processes not only saves the requirement on a carbon source, but also can save the energy source required by aeration when the phosphorus absorption is completed in the anoxic environment, and greatly reduces the amount of residual sludge. However, biological phosphorus removal of the existing sewage treatment system is finished by sludge discharge, the quantity of nitrifying bacteria is reduced while sludge discharge is carried out, and the nitrifying effect is poor, so that the risk that ammonia nitrogen in effluent does not reach standards exists while phosphorus removal is carried out, and meanwhile, competition of carbon sources exists between denitrifying bacteria and phosphorus accumulating bacteria.
CN2015167975. X discloses a denitrification synchronous denitrification dephosphorization bacteria enrichment domestication method. The method adopts a strategy of 'two-stage (anaerobic/aerobic first and anaerobic/anoxic second) enrichment and domestication, and a cyclic operation mode of water inlet, reaction, precipitation and drainage after high and low nutritional loads', the method enhances the enrichment quantity of dephosphorization microorganisms, improves the microorganism propagation rate, creates optimal conditions for microorganism propagation, and can enrich and domesticate denitrifying synchronous denitrification and dephosphorization bacteria under a low-temperature environment after anaerobic/aerobic operation for 40d and anaerobic/anoxic operation for 30d, wherein the phosphorus concentration in the effluent is stabilized at 0.5mg/L. However, the enriched flora of the method only has denitrifying phosphorus removal bacteria, and the application of the method in wastewater containing ammonia nitrogen pollutants for denitrification and phosphorus removal is limited.
CN200910082222.1 discloses a method for culturing aerobic granular sludge for simultaneously denitrifying and dephosphorizing domestic sewage at a low temperature. In the method, two sedimentation time modes are adopted, dissolved oxygen concentration DO, oxidation-reduction potential ORP and pH value are used as real-time control parameters in the biochemical reaction process, and stirring time and aeration time are controlled in real time, so that the aerobic granular sludge which simultaneously denitrifies and dephosphorizes at normal low temperature is obtained. The method cultures the thalli from the aspect of process control, and the quantity distribution, the overall activity and the adaptability of the thalli are still to be further improved.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a common enrichment method of nitrifying bacteria and denitrifying phosphorus accumulating bacteria. The nitrifying bacteria and the denitrifying phosphorus accumulating bacteria obtained by the method have high activity, good synergistic effect and strong adaptability, can be used in situ by on-site culture, and are particularly suitable for solving the problem that the total nitrogen and the total phosphorus in the existing sewage treatment field do not reach the standard.
The invention provides a method for jointly enriching nitrifying bacteria and denitrifying phosphorus accumulating bacteria, which comprises the following steps:
(1) Nitrifying bacteria culture stage: inoculating activated sludge containing nitrifying bacteria and denitrifying phosphorus accumulating bacteria, adding a culture solution, culturing nitrifying bacteria under aerobic aeration conditions, and stopping aeration and starting stirring when the ammonia nitrogen concentration in the culture solution is reduced to below 40%, preferably 10% -20%, of the initial concentration;
(2) And (3) culturing the denitrifying phosphorus accumulating bacteria: adding sugar esters into the culture solution obtained in the step (1), adding quaternary ammonium base after the dissolved oxygen is reduced to be less than 0.1mg/L, culturing denitrifying phosphorus accumulating bacteria, stopping stirring when the total phosphorus concentration in the culture solution is reduced to be less than 40%, preferably 10-20%, of the initial concentration, settling, and discharging supernatant to leave bacteria;
(3) And (3) taking the thalli obtained in the step (2) as activated sludge, repeating the processes of the step (1) and the step (2) until the relative abundance of nitrifying bacteria and denitrifying polyphosphoric bacteria in the cultured flora is more than 15% respectively, and ending the culture.
In the method, the ammonia nitrogen concentration in the culture solution used for the first time is 30-50mg/L, the COD concentration is 90-150mg/L, the total phosphorus concentration is 3-5mg/L, the ammonia nitrogen concentration in the culture solution used for the final time is 100-300mg/L, the COD concentration is 300-600mg/L, and the total phosphorus concentration is 10-30mg/L. The substrate concentration in the culture solution is gradually increased, the increasing amplitude of each time is 30-50mg/L according to ammonia nitrogen, COD is 90-150mg/L, and total phosphorus is 3-5mg/L. The conditions for increasing the substrate concentration of the culture solution are as follows: and (3) when the time for completing the step (1) and the step (2) by adopting the culture solution with the current substrate concentration is less than 60 percent (such as 40 to 60 percent) of the time for completing the step (1) and the step (2) by using the culture solution with the same substrate concentration for the first time, the substrate concentration in the culture solution used next time is increased.
In the method of the invention, the activated sludge containing nitrifying bacteria and denitrifying phosphorus accumulating bacteria can be contained in the raw sludge or added later. In the method, the activated sludge which is inoculated in the step (1) and is rich in nitrifying bacteria and denitrifying phosphorus accumulating bacteria is taken from sludge in a secondary sedimentation tank of a sewage treatment plant for treating sewage containing nitrogen and phosphorus, the inoculation amount is 2000-5000mg/L, and the relative abundance values of nitrifying bacteria and denitrifying phosphorus accumulating bacteria at the level are 0.1% -1.0% and 0.1% -1.0%, respectively.
In the method of the present invention, the free amino acid of step (1) comprises at least one of proline, hydroxyproline, glycine, arginine, etc., preferably proline. The amount of the free amino acid used in the step (1) is 0.001-1.0mg/L, preferably 0.01-0.10mg/L.
In the method of the invention, the culture conditions in the step (1) are as follows: the concentration of dissolved oxygen is 0.5-1.5mg/L, the pH is 7-8, and the temperature is 25-35 ℃.
In the method of the present invention, the sugar ester substance in the step (2) includes at least one of mouse Li Tangzhi, trehalose ester, sophorolipid, sucrose ester, etc., preferably rhamnose ester. The quaternary ammonium base is at least one of phosphorylcholine, betaine, tetramethylammonium hydroxide and the like, and preferably phosphorylcholine. The sugar ester substance in the step (2) is used in an amount of 0.001-1.0mg/L, preferably 0.01-0.10mg/L. The amount of the quaternary ammonium base is 0.001-1.0mg/L, preferably 0.01-0.10mg/L.
In the method of the invention, the culture conditions of the denitrifying phosphorus accumulating bacteria in the step (2) are as follows: the concentration of dissolved oxygen is 0-0.5mg/L, the pH is 7-8, and the temperature is 25-35 ℃.
In the method of the invention, the condition of the end of the culture is that the relative abundance of nitrifying bacteria and denitrifying polyphosphoric bacteria in the cultured flora is more than 15%, preferably 20% -45% and 20% -45% respectively.
The nitrifying bacteria and denitrifying phosphorus accumulating bacteria enriched by the method can be directly added into activated sludge of a sewage treatment field for use, and can also treat wastewater containing nitrogen and phosphorus in a proper biochemical reactor.
The method of the invention has the following advantages:
(1) In the nitrifying bacteria culture process, free amino acid is added, so that the effective transfer of oxygen and the full utilization of multiple matrixes can be improved, the competition of autotrophic nitrifying bacteria and heterotrophic nitrifying bacteria to the substrates is facilitated, the number of nitrifying bacteria is increased by the kinase activity, the nitrifying capacity is improved, meanwhile, the competition of phosphorus accumulating bacteria and nitrifying bacteria to dissolved oxygen can be reduced, and the aerobic phosphorus absorbing capacity of the phosphorus accumulating bacteria is improved.
(2) In the process of culturing denitrifying phosphorus accumulating bacteria, sugar ester substances and quaternary ammonium base are added in a matched manner, so that the activity of nitrate reductase can be improved, the utilization of nitrate as an electron acceptor by denitrifying bacteria is enhanced, and the growth and propagation of the denitrifying phosphorus accumulating bacteria are promoted.
(3) According to the invention, specific substances are added aiming at different culture stages, so that the synergistic growth of multiple bacterial groups can be promoted, the common enrichment of nitrifying bacteria and denitrifying phosphorus accumulating bacteria is realized, and the denitrification and phosphorus removing effects can be improved when the method is applied to a sewage treatment field.
Detailed Description
The technical scheme of the invention is further described in detail below with reference to examples. The embodiments and specific operation procedures are given on the premise of the technical scheme of the invention, but the protection scope of the invention is not limited to the following embodiments.
The experimental methods in the following examples, unless otherwise specified, are all conventional in the art. The experimental materials used in the examples described below were purchased from biochemical reagent stores unless otherwise specified.
In the method, the COD concentration is measured by GB11914-89 'determination of water quality chemical oxygen demand-dichromate method'; the ammonia nitrogen concentration is measured by using GB7478-87 method for measuring ammonium in water-distillation and titration, the total nitrogen concentration adopts GB 11894-89 water quality-total nitrogen determination-alkaline potassium persulfate digestion ultraviolet spectrophotometry, and the total phosphorus concentration adopts GB11893-89 ammonium molybdate spectrophotometry. The relative abundance at the nitrifying and denitrifying polyphosphazenes levels can be determined using high throughput sequencer analysis.
Example 1
(1) Taking the activated sludge of a secondary sedimentation tank of a sewage treatment plant, wherein the relative abundance values of nitrifying bacteria and denitrifying phosphorus accumulating bacteria are respectively 0.14% and 0.22%, and inoculating the activated sludge into a 5L organic glass reactor with aeration and stirring functions according to the sludge concentration (MLSS) of about 3000mg/L after inoculation to perform enrichment culture of thalli. The ammonia nitrogen concentration in the culture solution used for the first time is 30mg/L, the COD concentration is 90mg/L, the total phosphorus concentration is 3mg/L, and proline is added according to the concentration of 0.05 mg/L. Enrichment conditions of nitrifying bacteria are as follows: the concentration of dissolved oxygen is 1.0mg/L, the pH is 7.5, the temperature is 30 ℃, aeration is stopped when the concentration of ammonia nitrogen in the culture solution is reduced to 10% of the initial concentration, and stirring is started to enter the denitrification phosphorus accumulating bacteria culture stage.
(2) While stopping aeration and starting stirring, adding mouse Li Tangzhi according to the concentration of 0.05mg/L, adding phosphorylcholine according to the concentration of 0.05mg/L after the dissolved oxygen is reduced to less than 0.1mg/L, stopping stirring when the total phosphorus concentration is reduced to 20% of the initial concentration, settling for thirty minutes, and discharging the supernatant to leave the thallus at the bottom.
(3) And (3) taking the thalli obtained in the step (2) as activated sludge, and repeating the processes of the step (1) and the step (2). Wherein when the time for completing the step (1) and the step (2) by adopting the culture solution with the current substrate concentration is about 50% of the time for completing the step (1) and the step (2) by using the culture solution with the same substrate concentration for the first time, the substrate concentration in the culture solution used next time is increased. The substrate concentration in the culture solution is increased, the increasing amplitude of each time is 30mg/L according to the ammonia nitrogen, the COD is 90mg/L, and the total phosphorus is 3mg/L. Proline was added at a concentration of 0.05mg/L for each change of culture broth. The ammonia nitrogen concentration in the culture solution used for the last time is 150mg/L, the COD concentration is 450mg/L, and the total phosphorus concentration is 15mg/L. The relative abundance of nitrifying bacteria and denitrifying polyphosphoric bacteria in the final cultured flora respectively reaches 30% and 40%, and the culturing is finished and the thallus A is harvested for standby.
Example 2
The cultivation process and conditions were the same as in example 1 except that betaine was added at a concentration of 0.05mg/L after the dissolved oxygen was reduced to less than 0.1mg/L in the denitrifying phosphorus accumulating bacteria cultivation stage. The relative abundance of nitrifying bacteria and denitrifying polyphosphoric bacteria in the cultured flora after the culture is finished reaches 29% and 38%, respectively, and the thallus B is harvested for standby.
Example 3
The cultivation process and conditions were the same as in example 1, except that the substances added in the denitrification phosphorus accumulating bacteria cultivation stage and the dissolved oxygen stage were trehalose ester and betaine, respectively. The relative abundance of nitrifying bacteria and denitrifying polyphosphoric bacteria in the cultured flora after the culture is finished respectively reaches 27% and 36%, and the thallus C is harvested for standby.
Example 4
(1) Taking the activated sludge of a secondary sedimentation tank of a sewage treatment plant, wherein the relative abundance values of nitrifying bacteria and denitrifying phosphorus accumulating bacteria are respectively 0.36% and 0.47%, and inoculating the activated sludge into a 5L organic glass reactor with aeration and stirring functions according to the sludge concentration (MLSS) of about 4000mg/L after inoculation to perform enrichment culture of thalli. The ammonia nitrogen concentration in the culture solution used for the first time is 50mg/L, the COD concentration is 150mg/L, the total phosphorus concentration is 5mg/L, and hydroxyproline is added according to the concentration of 0.01 mg/L. Enrichment conditions of nitrifying bacteria are as follows: the concentration of dissolved oxygen is 1.0mg/L, the pH is 7.5, the temperature is 30 ℃, aeration is stopped when the concentration of ammonia nitrogen in the culture solution is reduced to 10% of the initial concentration, and stirring is started to enter the denitrification phosphorus accumulating bacteria culture stage.
(2) And stopping aeration and stirring, adding trehalose ester according to the concentration of 0.01mg/L, adding betaine according to the concentration of 0.05mg/L after the dissolved oxygen is reduced to less than 0.1mg/L, stopping stirring when the total phosphorus concentration is reduced to 10% of the initial concentration, settling for thirty minutes, and discharging the supernatant to leave the thallus at the bottom.
(3) And (3) taking the thalli obtained in the step (2) as activated sludge, and repeating the processes of the step (1) and the step (2). Wherein when the time for completing the step (1) and the step (2) by adopting the culture solution with the current substrate concentration is about 40% of the time for completing the step (1) and the step (2) by using the culture solution with the same substrate concentration for the first time, the substrate concentration in the culture solution used next time is increased. The substrate concentration in the culture solution is increased, the increasing amplitude of each time is 50mg/L according to the ammonia nitrogen, the COD is 100mg/L, and the total phosphorus is 5mg/L. Hydroxyproline was added at a concentration of 0.01mg/L for each replacement of the culture broth. The ammonia nitrogen concentration in the culture solution used for the last time is 300mg/L, the COD concentration is 550mg/L, and the total phosphorus concentration is 25mg/L. The relative abundance of nitrifying bacteria and denitrifying polyphosphoric bacteria in the final cultured flora respectively reaches 25% and 34%, and the culturing is finished and the thallus D is harvested for standby.
Example 5
(1) Taking active sludge in a secondary sedimentation tank of a sewage treatment plant, wherein relative abundance values of nitrifying bacteria and denitrifying phosphorus accumulating bacteria are respectively 0.36% and 0.47%, and inoculating the active sludge into a 5L organic glass reactor with aeration and stirring functions according to the sludge concentration (MLSS) of about 3500mg/L after inoculation to perform enrichment culture of thalli. The ammonia nitrogen concentration in the culture solution used for the first time is 40mg/L, the COD concentration is 120mg/L, the total phosphorus concentration is 4mg/L, and arginine is added according to the concentration of 0.1 mg/L. Enrichment conditions of nitrifying bacteria are as follows: the concentration of dissolved oxygen is 1.0mg/L, the pH is 7.5, the temperature is 30 ℃, aeration is stopped when the concentration of ammonia nitrogen in the culture solution is reduced to 10% of the initial concentration, and stirring is started to enter the denitrification phosphorus accumulating bacteria culture stage.
(2) While stopping aeration and starting stirring, sucrose ester is added according to the concentration of 0.05mg/L, tetramethylammonium hydroxide is added according to the concentration of 0.01mg/L after the dissolved oxygen is reduced to less than 0.1mg/L, stirring is stopped when the total phosphorus concentration is reduced to 15% of the initial concentration, and the supernatant is discharged after thirty minutes of sedimentation, so that the bottom thalli are left.
(3) And (3) taking the thalli obtained in the step (2) as activated sludge, and repeating the processes of the step (1) and the step (2). Wherein when the time for completing the step (1) and the step (2) by adopting the culture solution with the current substrate concentration is about 60% of the time for completing the step (1) and the step (2) by using the culture solution with the same substrate concentration for the first time, the substrate concentration in the culture solution used next time is increased. The substrate concentration in the culture solution is increased, the increasing amplitude of each time is 40mg/L according to the ammonia nitrogen, the COD is 100mg/L, and the total phosphorus is 5mg/L. Arginine was added at a concentration of 0.1mg/L for each change of culture broth. The ammonia nitrogen concentration in the culture solution used for the last time is 200mg/L, the COD concentration is 500mg/L, and the total phosphorus concentration is 25mg/L. The relative abundance of nitrifying bacteria and denitrifying polyphosphoric bacteria in the final cultured flora respectively reaches 24% and 36%, and the culturing is finished and the thallus E is harvested for standby.
Comparative example 1
In comparison with example 1, except that proline added at 0.05mg/L was changed to betaine in the nitrifying bacteria enrichment stage of step (1), and in the case of the same total culture time as in example 1, the cells F were harvested, wherein the relative abundance at the nitrifying bacteria and denitrifying polyphosphbacteria levels reached 16% and 12%, respectively.
Comparative example 2
In comparison with example 1, except that the mice Li Tangzhi and phosphorylcholine were not added in step (2), and in the case where the total cultivation time was the same as in example 1, the cells G were harvested, wherein the relative abundance at the nitrifying bacteria and denitrifying polyphosphbacteria levels reached 18% and 10%, respectively.
Comparative example 3
In comparison with example 1, except that phosphorylcholine was added simultaneously with rhamnose ester in step (2), bacterial cells H were harvested in which the relative abundance at the nitrifying bacteria and denitrifying polyphosphazenes levels reached 17% and 16%, respectively, with the same total cultivation time as in example 1.
Comparative example 4
In comparison with example 1, except that the substrate concentration in the culture broth was not increased during the culture, and in the case where the total culture time was the same as in example 1, the cells I were harvested, in which the relative abundance amounts at the nitrifying bacteria and denitrifying polyphosphbacteria levels reached 21% and 18%, respectively.
Effect verification
And 10 reactors with the effective volumes of 5L are taken in a laboratory to verify the denitrification and dephosphorization effects of the thalli. The sewage water quality for experiments is ammonia nitrogen concentration 100mg/L, total nitrogen concentration 110mg/L, COD concentration 300mg/L and total phosphorus concentration 8mg/L. The reactor was inoculated with activated sludge at a sludge concentration of 2000mg/L, and then the cells obtained in the examples and comparative examples were inoculated into the respective reactors at the same inoculum size. The SBR process is adopted for treatment, the dissolved oxygen in the stirring stage is controlled at 0.5mg/L, the dissolved oxygen in the aeration stage is controlled at 3mg/L, the pH is 7.8, the temperature is 32 ℃, the reaction is ended after 24 hours, the concentration of pollutants in the supernatant fluid in each reactor is sampled and analyzed, and the specific results are shown in Table 1.
TABLE 1 effects of treatment on various bacterial cells
| Thallus | COD of effluent, mg/L | Effluent NH 3 -N，mg/L | Total nitrogen in effluent, mg/L | Total phosphorus in effluent, mg/L |
| Thallus A | 43.2 | 7.9 | 25.1 | 0.49 |
| Thallus B | 48.6 | 8.2 | 27.4 | 0.56 |
| Thallus C | 49.1 | 9.4 | 28.3 | 0.53 |
| Thallus D | 52.3 | 11.3 | 29.4 | 0.67 |
| Thallus E | 54.4 | 10.7 | 30.1 | 0.71 |
| Thallus F | 51.8 | 12.1 | 37.9 | 4.96 |
| Thallus G | 55.7 | 13.6 | 38.8 | 4.81 |
| Thallus H | 57.3 | 14.2 | 41.8 | 3.93 |
| Thallus I | 62.6 | 18.3 | 47.0 | 4.89 |
| Control without adding bacteria | 75.2 | 21.4 | 56.7 | 7.12 |
As can be seen from the data in Table 1, the COD concentration of the treated effluent is lower than 55mg/L, the ammonia nitrogen concentration is lower than 12mg/L, the total nitrogen concentration is lower than 35mg/L, and the total phosphorus concentration is lower than 1.0mg/L by using the bacterial cells cultured by the invention, so that the biological treatment effect of sewage is obviously improved, and the simultaneous denitrification and dephosphorization are realized.
Claims (16)
1. A method for jointly enriching nitrifying bacteria and denitrifying phosphorus accumulating bacteria comprises the following steps:
(1) Nitrifying bacteria culture stage: inoculating activated sludge containing nitrifying bacteria and denitrifying phosphorus accumulating bacteria, adding a culture solution, culturing nitrifying bacteria under aerobic aeration condition, stopping aeration and starting stirring when ammonia nitrogen concentration in the culture solution is reduced to below 40% of the initial concentration;
(2) And (3) culturing the denitrifying phosphorus accumulating bacteria: stopping aeration, starting stirring, adding sugar ester substances, adding quaternary ammonium base when dissolved oxygen is reduced to less than 0.1mg/L, culturing denitrifying phosphorus accumulating bacteria, stopping stirring when the total phosphorus concentration in the culture solution is reduced to less than 40% of the initial concentration, settling, and discharging supernatant to leave thalli;
(3) And (3) taking the thalli obtained in the step (2) as activated sludge, repeating the processes of the step (1) and the step (2) until the relative abundance of nitrifying bacteria and denitrifying polyphosphoric bacteria in the cultured flora is more than 15% respectively, and ending the culture.
2. A method according to claim 1, characterized in that: and (1) stopping aeration and starting stirring when the ammonia nitrogen concentration in the culture solution is reduced to 10% -20% of the initial concentration.
3. A method according to claim 1, characterized in that: and (2) stopping stirring when the total phosphorus concentration in the culture solution is reduced to 10-20% of the initial concentration, settling, and discharging the supernatant to leave thalli.
4. A method according to claim 1, characterized in that: the concentration of ammonia nitrogen in the culture solution used for the first time is 30-50mg/L, the concentration of COD is 90-150mg/L, the concentration of total phosphorus is 3-5mg/L, the substrate concentration in the culture solution adopted in the process of repeating the step (1) and the step (2) is gradually increased, the increasing amplitude of each time is 30-50mg/L according to the ammonia nitrogen, the COD is 90-150mg/L, and the total phosphorus is 3-5mg/L; the ammonia nitrogen concentration in the final used culture solution is 100-300mg/L, the COD concentration is 300-600mg/L, and the total phosphorus concentration is 10-30mg/L.
5. A method according to claim 1 or 4, characterized in that: the conditions for increasing the substrate concentration of the culture solution are as follows: and (3) when the time for completing the step (1) and the step (2) by adopting the culture solution with the current substrate concentration is less than 60% of the time for completing the step (1) and the step (2) by using the culture solution with the same substrate concentration for the first time, the substrate concentration in the culture solution used next time is increased.
6. A method according to claim 1, characterized in that: the activated sludge which is inoculated in the step (1) and is rich in nitrifying bacteria and denitrifying phosphorus accumulating bacteria is taken from sludge in a secondary sedimentation tank of a sewage treatment plant for treating sewage containing nitrogen and phosphorus, the inoculum size is 2000-5000mg/L, and the relative abundance values of nitrifying bacteria and denitrifying phosphorus accumulating bacteria at the level are 0.1% -1.0% and 0.1% -1.0%, respectively.
7. A method according to claim 1, characterized in that: the free amino acid in the step (1) comprises at least one of proline, hydroxyproline, glycine and arginine; the free amino acid in the step (1) is used in an amount of 0.001-1.0mg/L.
8. The method of claim 7, wherein: the free amino acid in step (1) is proline.
9. The method of claim 7, wherein: the free amino acid in the step (1) is used in an amount of 0.01-0.10mg/L.
10. A method according to claim 1, characterized in that: the culture conditions in the step (1) are as follows: the concentration of dissolved oxygen is 0.5-1.5mg/L, the pH is 7-8, and the temperature is 25-35 ℃.
11. A method according to claim 1, characterized in that: the sugar ester substance in the step (2) comprises at least one of mouse Li Tangzhi, trehalose lipid, sophorolipid and sucrose ester; the quaternary ammonium base is at least one of phosphorylcholine, betaine and tetramethylammonium hydroxide.
12. The method of claim 11, wherein: the sugar ester substance in the step (2) is mouse Li Tangzhi; the quaternary ammonium base is phosphorylcholine.
13. A method according to claim 1 or 11, characterized in that: the dosage of the sugar ester substance in the step (2) is 0.001-1.0mg/L; the dosage of the quaternary ammonium base is 0.001-1.0mg/L.
14. The method of claim 13, wherein: the dosage of the sugar ester substance in the step (2) is 0.01-0.10mg/L; the dosage of the quaternary ammonium base is 0.01-0.10mg/L.
15. A method according to claim 1, characterized in that: the culture conditions of the denitrifying phosphorus accumulating bacteria in the step (2) are as follows: the concentration of dissolved oxygen is 0-0.5mg/L, the pH is 7-8, and the temperature is 25-35 ℃.
16. A method according to claim 1, characterized in that: the condition of the end of the culture is that the relative abundance of nitrifying bacteria and denitrifying polyphosphoric bacteria in the cultured flora reaches 20% -45% and 20% -45% respectively.
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