CN115627246A - Application of aerobic denitrifying strain ZW-2 in ammonia nitrogen sewage treatment - Google Patents
Application of aerobic denitrifying strain ZW-2 in ammonia nitrogen sewage treatment Download PDFInfo
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Abstract
The invention provides an application of an aerobic denitrifying strain ZW-2 in ammonia nitrogen sewage treatment, wherein the strain ZW-2 is Pseudomonas stutzeri, and is preserved in the general microbiological culture Collection center of the Committee for culture Collection of microorganisms in 2022, 9 months and 19 days, and the preservation number is CGMCC NO.25760. The aerobic denitrifying bacteria ZW-2 and the nitrifying bacteria are combined, so that the aim of converting inorganic nitrogen in a water body into nitrogen-containing gas by the coupling of nitrification and aerobic denitrification is really fulfilled, the TN removal rate is up to more than 80%, and the effluent meets the first-level A standard.
Description
Technical Field
The invention relates to the technical field of sewage treatment, in particular to application of an aerobic denitrification strain ZW-2 in ammonia nitrogen sewage treatment.
Background
The traditional sewage treatment process generally adopts an A2O process flow, and mainly relies on the combination of aerobic nitrobacteria and anaerobic denitrifying bacteria to denitrify and remove ammonia from sewage. However, because of the great difference in growth environment, physiological and biochemical (especially dissolved oxygen) and the like, the conventional aerobic nitrifying bacteria and anaerobic denitrifying bacteria are always required to independently operate in 2 reactors, which causes the defects of long time consumption, low efficiency, high cost and the like of the conventional biological denitrification technology, and obviously increases the construction and operation costs. In addition, in the A2O process, the mixed liquid (nitrification liquid) in the O tank needs to flow back to the a tank (anoxic tank) to realize denitrification process for denitrification, which greatly increases process complexity and operation cost.
Traditionally, microorganisms have been able to remove nitrate from water only by reducing it anaerobically to produce gaseous products. This anaerobic process can be divided into heterotrophic denitrification and autotrophic denitrification depending on the type of electron donor and carbon source used by the microorganism. Heterotrophic denitrification is characterized in that facultative anaerobes reduce nitrate by using organic matters as electron donors and carbon sources, while autotrophic denitrification reduces nitrate to nitrogen-containing compounds such as nitrogen by using inorganic carbon as a carbon source and using reduced sulfur, ferrous iron, hydrogen and the like as electron donors. However, natural water bodies, especially surface water, usually contain high dissolved oxygen (5-10 mg/L) and cannot create sufficient anaerobic environment for traditional denitrification to remove nitrate. In addition, the conventional nitrification-denitrification microorganisms have long generation propagation time and low nitrification-denitrification rate, and cannot tolerate the conditions of high ammonia, high COD, extreme environments (pH, temperature, heavy metal) and the like, so that the application range of the microorganisms is limited.
Disclosure of Invention
In view of the defects of the prior art, the invention provides the aerobic denitrifying bacteria for efficiently treating the nitrogen-containing sewage, the strain is subjected to pre-membrane formation, the enriched efficient nitrifying bacteria liquid is also subjected to pre-membrane formation, after the two efficient nitrifying bacteria liquid are successfully subjected to membrane formation, two membrane formation fillers are mixed and combined with an MBBR (moving bed biofilm reactor) process to simulate the treatment of the wastewater, and the result shows that the denitrification and ammonia removal capacity is high, COD (chemical oxygen demand) can be degraded, the effluent quality is stable, and the effluent quality meets the first-level A standard.
The technical scheme of the invention mainly comprises the following contents:
the invention relates to an aerobic denitrifying strain ZW-2 which is identified to be Pseudomonas stutzeri (Pseudomonas stutzeri). The strain has been preserved in China general microbiological culture Collection center (CGMCC for short, with the address of No. 3 Xilu No. 1 Beijing, chaoyang district, beijing) with the preservation number of CGMCC NO.25760 at 9 months and 19 days of 2022.
In another aspect, the invention relates to the application of the aerobic denitrifying strain ZW-2 in ammonia nitrogen sewage treatment.
Preferably, the applying step comprises: filling the filler of the strain ZW-2 biofilm formation into a sewage treatment device, adding flowing ammonia nitrogen sewage into the sewage treatment device, carrying out sewage treatment under an aerobic condition, and detecting the quality of the effluent until the quality meets the requirements.
Preferably, the applying step comprises: filling the filler of the biofilm formation of the nitrifying bacteria liquid and the filler of the biofilm formation of the strain ZW-2 in a sewage treatment device, adding flowing ammonia nitrogen sewage into the sewage treatment device, carrying out sewage treatment under aerobic condition, and detecting the effluent quality until the effluent quality meets the requirements.
Preferably, the volume ratio of the filler of the nitrifying bacteria liquid biofilm formation to the filler of the bacterial strain ZW-2 biofilm formation is (4-5) to (4-5).
Preferably, the filling rate of the filler in the sewage treatment plant is at least 90%.
Preferably, the filler is a polyurethane filler.
Preferably, the filler of the biofilm formation of the nitrifying bacteria liquid and the filler of the biofilm formation of the strain ZW-2 are synchronously or step-by-step filled in a sewage treatment device.
Preferably, the nitrifying bacteria are obtained by the following method: adding a sewage sample into a culture medium, performing shake culture, measuring the concentrations of ammonia nitrogen and nitrite nitrogen every day, lasting for at least 5 days, respectively calculating the ammonia nitrogen consumption speed and the nitrite nitrogen accumulation speed, and identifying a group with an acceleration effect; and (4) further subculturing the group with the best effect, wherein the steps are the same as the above, and each round of cultivation lasts for 5-6 days for at least 4 rounds, so that the nitrifying bacteria liquid is finally obtained.
Preferably, the conditions for shake culture are: the temperature is 27 ℃, and the rotating speed is 150r/min.
Preferably, the culture medium is: adding MgSO 2 per liter of water 4 .7H 2 O 0.2g，K 2 HPO 4 0.1g，FeCl 3 0.1g，Fe-EDTA 0.4g，NaHCO 3 1.0g，CaCl 2 .2H 2 O2.0g, trace elements 1ml, (NH) 4 ) 2 SO 4 0.48g。
The invention has the following effects:
(1) The utilization of aerobic denitrifying microorganisms makes the synchronous nitrification and denitrification possible, so that the capital investment cost is greatly saved, the alkalinity generated in the denitrification process can also compensate the acidity consumed by nitrification, and the operation cost is further reduced.
(2) Compared with the traditional anaerobic denitrifying bacteria, the aerobic denitrifying bacteria ZW-2 have the advantages of fast growth, easy propagation, easy control, strong environmental adaptability and the like, so that the sewage treatment efficiency is greatly improved, and the effluent is more stable compared with the traditional process.
(3) The aerobic denitrification microbial agent biofilm formation is used for treating sewage, so that the starting time of the biofilm reactor can be obviously shortened, and the running cost and the time cost are reduced.
Drawings
FIG. 1: pseudomonas stutzeri morphogram.
FIG. 2: a graph of the denitrification characteristic analysis result of Pseudomonas stutzeri ZW-2.
FIG. 3: example 3 screening of culture conditions for Strain ZW-2 results (rotational speed).
FIG. 4: example 3 screening of culture conditions for Strain ZW-2 results (carbon to nitrogen ratio).
FIG. 5 is a schematic view of: example 3 culture conditions of Strain ZW-2 results of screening experiments (carbon source).
FIG. 6: EXAMPLE 3 culture conditions of Strain ZW-2 the results of the experiment were screened (temperature).
FIG. 7: example 3 culture conditions of Strain ZW-2 results of screening experiments (initial pH).
FIG. 8: the sewage treatment device is a schematic structural diagram.
FIG. 9: example 4 application result statistical chart (ammonia nitrogen concentration) of the strain ZW-2 in ammonia nitrogen sewage treatment; wherein, stage 1: nitrifying bacteria liquid; and (2) stage: nitrifying bacteria solution and aerobic denitrifying bacteria (ZW-2).
FIG. 10: example 4 statistical chart of application results of strain ZW-2 in Ammonia nitrogen wastewater treatment (NO) 3 -N concentration)
FIG. 11: example 4 statistical graph of application results of strain ZW-2 in Ammonia nitrogen wastewater treatment (NO) 2 -N concentration)
FIG. 12: example 4 application result statistical chart of Strain ZW-2 in Ammonia nitrogen wastewater treatment (TN concentration)
FIG. 13: example 4 statistical chart of application results (COD concentration) of Strain ZW-2 in Ammonia-nitrogen wastewater treatment
Detailed Description
In order to better understand the technical content of the invention, specific examples are provided below to further illustrate the invention.
Example 1 isolation and purification of Pseudomonas stutzeri ZW-2
1.1 culture Medium
Aerobic denitrification culture medium: 2g/L NaCl, KH 2 PO 4 1.5g/L，MgSO 4 ·7H 2 O 0.01g/L，Na 2 HPO 4 7.87g/L, 3.375g/L sodium succinate, naNO 3 0.605g/L, and 2ml/L of trace element solution.
Separating and purifying the culture medium: naCl 2g/L, KH 2 PO 4 1.5g/L，MgSO 4 ·7H 2 O 0.01g/L，Na 2 HPO 4 7.87g/L, 3.375g/L sodium succinate, naNO 3 0.605g/L, 2ml/L of trace element solution and 20g/L of agar powder.
Solution of trace elements: EDTA 50g/L, feSO 4 ·7H 2 O 5.0g/L，CaCl 2 5.5g/L，CuSO 4 ·5H 2 O 1.57g/L，ZnSO 4 2.2g/L，MnCl 2 ·4H 2 O 5.06g/L，CoCl 2 ·6H 2 O 1.61g/L，(NH 4 ) 6 Mo 7 O 2 ·4H 2 O 1.1g/L。
The above culture medium is prepared, and then sterilized at 121 deg.C under high pressure for 20min.
1.2 enrichment, isolation and screening of the strains
Taking 10mL of activated sludge sample, placing the sample in a 250mL conical flask filled with 90mL of sterile aerobic denitrification culture medium, culturing for 2d at constant temperature of 30 ℃ at 150r/min, taking 10mL of culture solution, transferring the culture solution into a new 90mL of aerobic denitrification culture medium, continuously culturing for 3d under the same condition, and repeating for 3 times. The enriched solution is subjected to gradient dilution (10) using a phosphoric acid buffer solution -2 ～10 -7 ) 0.1mL of each gradient was plated on solid isolation and purification medium and cultured in an inverted state in an incubator at 30 ℃ until a single colony was evident. And selecting colonies with different forms, and performing plate streaking culture for multiple times to finally obtain the purified strain ZW-2. The strain morphology is characterized as follows: the colony is round, wrinkled and yellow, and the thallus is short rod-shaped and 5-6 μm long (FIG. 1). The strain is sent to the China general microbiological culture Collection center for preservation, and the preservation number is CGMCC NO.25760.
Example 2 Pseudomonas stutzeri ZW-2 Denitrification characterization
Under aerobic conditionsNext, the strain ZW-2 was inoculated to 100mg/L NO 3 In a medium with-N as the sole nitrogen source, after 32h of cultivation, the strain is resistant to NO 3 The removal rate of-N reaches 97.82 percent, wherein the TN removed by aerobic denitrification is 58.7 percent, and a small amount of NO is generated when the culture is carried out for 16h 2 N accumulated but disappeared completely after 24 h. The results show that the strain ZW-2 has stronger aerobic denitrification capability. (FIG. 2)
EXAMPLE 3 Strain ZW-2 culture Condition screening
The research shows that the denitrification conditions of the strain ZW-2 are as follows: the carbon source is sodium succinate or glucose, C/N = 8-25, dissolved oxygen (rotation speed) 0-150 r/min, temperature 30-37 ℃, pH = 7-10. Wherein, the optimal denitrification conditions are as follows: the carbon source is sodium succinate, C/N =15, dissolved oxygen (rotation speed) 100r/min, temperature 37 ℃, pH =9. (FIGS. 3 to 7)
Example 4 application of Strain ZW-2 in Ammonia nitrogen wastewater treatment
4.1 enrichment of nitrifying bacteria liquid
4.1.1 enrichment Medium:
MgSO 4 .7H 2 O 0.2g，K 2 HPO 4 0.1g，FeCl 3 0.1g，Fe-EDTA 0.4g，NaHCO 3 1.0g，CaCl 2 .2H 2 o2.0g, trace elements 1ml, (NH) 4 ) 2 SO 4 0.48g, 1000ml of distilled water, pH 7.2.
Trace elements: cuSO 4 5H 2 O 0.075g，ZnSO 4 7H 2 O 0.3g，CoCl 2 .6H 2 O 0.375g，MnCl 2 .2H 2 O 0.3g，EDTA 0.5g，NaMoO 4 2H 2 O 0.22g，H 3 BO 4 0.014g, 1000ml of distilled water.
4.1.2 enrichment method
Taking 5ml of water sample into a sterilized 500ml conical flask, adding 200ml of culture medium, performing shake culture at 27 ℃ and 150r/min, measuring the concentration of ammonia nitrogen and nitrite nitrogen every day, continuing for 5 days, respectively calculating the ammonia nitrogen consumption speed and the nitrite nitrogen accumulation speed (mg/L/d), and identifying groups with acceleration effect. And (4) carrying out subculture on the group with the best effect, wherein the steps are the same as the above, and each round of culture is carried out for 5-6 days and is continued for 4 rounds, so as to finally obtain the nitrifying bacteria liquid with high-efficiency ammonia removal. The microbial composition in the nitrifying bacteria liquid is as follows: nitrosomonas, nitrosococcus, pseudomonas, nitrospirillum, and the like.
4.2 treatment of wastewater
The sewage treatment device is shown in figure 8.
The experimental process comprises the following steps: firstly filling a successful filler (the filler is a polyurethane filler) for nitrifying bacteria biofilm formation in a sewage treatment device, wherein the filling volume is about 50 percent of the device, adding sewage containing ammonia nitrogen into the sewage treatment device for a flow experiment, wherein the hydraulic retention time of the sewage in the device is 24 hours, the temperature is natural water temperature, continuous aeration is kept, the dissolved oxygen in the water is 3-5 mg/L, and the sewage treatment effect is shown as stage 1 in fig. 9-13. After 30 days of treatment, the fillers (polyurethane fillers) successfully formed by the aerobic denitrifying bacteria ZW-2 in a biofilm mode are continuously placed in the sewage treatment device, the filling volume is about 50% of the device, the total filling rate of the two fillers is more than 90%, the flowing experiment is continuously carried out, and the sewage treatment effect is shown as stage 2 in fig. 9-13.
The bacteria liquid biofilm culturing judgment method comprises the following steps: and (3) respectively using the two bacteria liquids to carry out filler biofilm culturing treatment, wherein the nitrifying bacteria biofilm culturing lasts for about 3 weeks, the denitrifying bacteria biofilm culturing lasts for about 2 weeks (10-14 days), the effluent quality is detected during the biofilm culturing treatment period, and successful biofilm culturing is represented when the effluent quality is stable and yellow biofilms are uniformly distributed on the surface of a carrier under the observation of naked eyes.
4.3 results of the experiment
The results are shown in FIGS. 9 to 13. In the figure, the stage 1 represents the water treatment result before the nitrobacteria biofilm culturing filler is added but the aerobic denitrifying bacteria ZW-2 biofilm culturing filler is not added, and the stage 2 represents the result after the aerobic denitrifying bacteria ZW-2 biofilm culturing filler is added. The result shows that the ammonia nitrogen is coupled by nitrification and aerobic denitrification through the combination of aerobic denitrifying bacteria and nitrifying bacteria, so that the aim of denitrification for converting inorganic nitrogen in a water body into nitrogen-containing gas is really fulfilled (the total nitrogen is not removed by assimilation and absorption of the aerobic denitrifying bacteria), the TN removal rate is up to more than 80%, and the effluent meets the first-level A standard.
The above description is only exemplary of the present invention and should not be taken as limiting the invention, as any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. The application of the aerobic denitrifying strain ZW-2 in ammonia nitrogen sewage treatment is characterized in that the strain ZW-2 is Pseudomonas stutzeri, and the strain is preserved in the general microbiological culture collection center of the microbiological culture collection management committee in 2022, 9 months and 19 months, and the preservation number is CGMCC NO.25760.
2. The use of claim 1, wherein the applying step comprises: filling the filler of the strain ZW-2 biofilm formation in a sewage treatment device, adding flowing ammonia nitrogen sewage into the sewage treatment device, carrying out sewage treatment under an aerobic condition, and detecting the quality of the effluent until the effluent meets the requirement.
3. The use according to claim 2, wherein the applying step comprises: filling the filler of the biofilm formation of the nitrifying bacteria liquid and the filler of the biofilm formation of the strain ZW-2 into a sewage treatment device, adding flowing ammonia nitrogen sewage into the sewage treatment device, carrying out sewage treatment under an aerobic condition, and detecting the quality of effluent until the effluent meets the requirements.
4. The application of claim 3, wherein the volume ratio of the filler of the nitrifying bacteria liquid biofilm formation to the filler of the bacterial strain ZW-2 biofilm formation is (4-5) to (4-5).
5. Use according to claim 2, wherein the filling rate of the filler in the sewage treatment plant is at least 90%.
6. Use according to claim 1, characterized in that the filler is a polyurethane filler.
7. The application of claim 1, wherein the filler for biofilm culturing of nitrifying bacteria liquid and the filler for biofilm culturing of the strain ZW-2 are filled in a sewage treatment device synchronously or step by step.
8. The use according to claim 1, wherein the nitrifying bacteria are obtained by: adding a sewage sample into a culture medium, performing shake culture, measuring the concentrations of ammonia nitrogen and nitrite nitrogen every day, lasting for at least 5 days, respectively calculating the ammonia nitrogen consumption speed and the nitrite nitrogen accumulation speed, and identifying groups with acceleration effects; and (4) further subculturing the group with the best effect, wherein the steps are the same as the above, and each round of cultivation lasts for 5-6 days for at least 4 rounds, so that the nitrifying bacteria liquid is finally obtained.
9. The use of claim 8, wherein the conditions for shake culture are: the temperature is 27 ℃, and the rotating speed is 150r/min.
10. Use according to claim 8, wherein the culture medium is: mgSO is added per liter of water 4 .7H 2 O 0.2g，K 2 HPO 4 0.1g，FeCl 3 0.1g，Fe-EDTA 0.4g，NaHCO 3 1.0g，CaCl 2 .2H 2 O2.0g, trace elements 1ml, (NH) 4 ) 2 SO 4 0.48g。
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| CN117721031A (en) * | 2023-05-11 | 2024-03-19 | 恒臻(无锡)生物科技有限公司 | Ammonia nitrogen degrading bacteria and application thereof in leather wastewater treatment |
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