CN114231441A - Ammonia oxidizing bacteria agent applied to sewage ammonia nitrogen treatment and preparation method thereof - Google Patents

Ammonia oxidizing bacteria agent applied to sewage ammonia nitrogen treatment and preparation method thereof Download PDF

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Publication number
CN114231441A
CN114231441A CN202111425619.3A CN202111425619A CN114231441A CN 114231441 A CN114231441 A CN 114231441A CN 202111425619 A CN202111425619 A CN 202111425619A CN 114231441 A CN114231441 A CN 114231441A
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ammonia oxidizing
oxidizing bacteria
sewage
culture medium
ammonia
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金文标
陈洪一
韩炜
车林
毛新瑞
张乃心
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Shenzhen Graduate School Harbin Institute of Technology
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Shenzhen Graduate School Harbin Institute of Technology
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • 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
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C02F3/348—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00—Nature of the contaminant
    • C02F2101/10—Inorganic compounds
    • C02F2101/16—Nitrogen compounds, e.g. ammonia

Abstract

The invention provides an ammonia oxidizing bacteria microbial inoculum applied to sewage ammonia nitrogen treatment and a preparation method thereof, wherein the microbial inoculum comprises one or more aerobic ammonia oxidizing bacteria, and the ammonia oxidizing bacteria microbial inoculum provided by the invention can have a better removal effect in sewage with a low carbon-nitrogen ratio, reduce the concentration of total nitrogen effluent, and achieve a total nitrogen removal rate of about 70%; the invention provides technical support for improving the nitrogen emission standard of the domestic sewage treated by the biological method, and has positive significance for improving the effluent standard of the domestic sewage.

Description

Ammonia oxidizing bacteria agent applied to sewage ammonia nitrogen treatment and preparation method thereof
Technical Field
The invention relates to the technical field of environmental microorganisms, in particular to an ammonia oxidizing bacterium agent applied to sewage ammonia nitrogen treatment and a preparation method thereof.
Background
There are two main approaches to biological denitrification of wastewater, one is through nitrification/denitrification process and the other is through anaerobic ammonia oxidation process. The nitrification reaction is a process of converting ammonia nitrogen into nitrite nitrogen or nitrate nitrogen, wherein ammonia nitrogen is converted into nitrite by ammonia oxidizing microorganisms under aerobic conditions and then oxidized into nitrate by nitrite oxidizing bacteria, wherein the ammonia oxidation reaction is the rate-limiting step in the process. The microorganisms capable of realizing the ammonia oxidation process are divided according to the metabolic types and can be divided into autotrophic nitrifying bacteria and heterotrophic nitrifying bacteria.
Domestic sewage in China generally faces the problem of low carbon and nitrogen, and the shortage of carbon sources causes the traditional sewage treatment process (such as A/O, A)2O, etc.) is difficult to reach the standard, especially the emission standard of total nitrogen is improved year by year. In the biological denitrification technology, heterotrophic nitrifiers can intensify competition among different strains on a carbon source, and have adverse effects on the subsequent denitrification and phosphorus absorption processes; autotrophic nitrifying bacteria are the research focus of the current sewage denitrification, and comprise ammonia oxidizing bacteria, ammonia oxidizing archaea, anaerobic ammonia oxidizing bacteria and the like, but the enrichment and separation of the ammonia oxidizing bacteria and the anaerobic ammonia oxidizing bacteria are very difficult, and the effect of improving the total nitrogen removal rate is difficult to achieve.
Disclosure of Invention
In order to solve the problems, the invention provides an ammonia oxidizing bacteria agent applied to sewage ammonia nitrogen treatment and a preparation method thereof. The specific contents are as follows:
in a first aspect, the invention relates to an ammonia oxidizing bacteria microbial inoculum for treating ammonia nitrogen in domestic sewage, which comprises one or more aerobic ammonia oxidizing bacteria.
Preferably, the aerobic ammonia oxidizing bacteria are gram-positive bordetella.
Preferably, the concentration of the microbial inoculum is 108-109one/mL.
Preferably, the aerobic ammonia oxidizing bacteria have positive reactions of contact enzyme and oxidase; starch hydrolysis, gelatin liquefaction, methyl red reaction, V-P reaction, and citrate utilization reaction were all negative.
In a second aspect, a method for preparing an ammonia oxidizing bacteria agent for ammonia nitrogen treatment of domestic sewage comprises the following steps: inoculating one or more aerobic ammonia oxidizing strains into a culture medium, and culturing at 30 ℃ and 120r/min for 24h to obtain an ammonia oxidizing bacterium agent.
Preferably, the raw material of the culture medium is sterilized sewage, and the culture medium further comprises beef extract, trace elements, magnesium elements and potassium elements.
Preferably, the concentrations of the beef extract, the trace elements, the magnesium elements and the potassium elements in the culture medium are 5g/L, 0.5mL/L, 1.0mg/L and 1.5mg/L respectively.
Preferably, the one or more aerobic ammonia oxidizing strains are selected by:
step 1: standing the sludge-water mixture taken out of the aeration tank, and removing supernatant to obtain sludge;
step 2: taking part of the sludge in a centrifugal tube, diluting with sterile normal saline and fully oscillating;
and step 3: taking a preset milliliter of the oscillated liquid to a conical flask filled with a domestication culture medium, and placing the conical flask in a constant-temperature shaking table to culture in a dark place; wherein the rotating speed of the constant-temperature shaking table is a preset rotating speed;
and 4, step 4: measuring the concentration of ammoniacal nitrogen in the culture solution in the constant-temperature shaking table at intervals of preset time, when the concentration of ammoniacal nitrogen is lower than a preset concentration value, taking the preset milliliter of culture solution, transferring the preset milliliter of culture solution into a fresh domestication culture medium, and repeating the step 4;
and 5: and (4) after repeating the step (4) for preset times, taking the culture solution to perform gradient dilution, flat plate coating and streak separation so as to screen out one or more aerobic ammonia oxidizing strains.
Compared with the prior art, the invention has the following beneficial effects:
the invention provides a microbial inoculum of ammonia oxidizing bacteria, which comprises one or more aerobic ammonia oxidizing bacteria, wherein the aerobic ammonia oxidizing bacteria are autotrophic denitrogenation bacteria, and ammonia nitrogen in sewage is used as a main nutrient source instead of a common carbon source, so that the demand of the aerobic ammonia oxidizing bacteria on the carbon source is reduced, in the sewage with a low carbon-nitrogen ratio, a good ammonia nitrogen removal effect can be achieved, meanwhile, the competition of different strains on the carbon source under the condition of low carbon-nitrogen ratio is avoided, and the activity of effective bacteria is improved; meanwhile, the strain obtained by the embodiment of the invention is convenient to screen, and simultaneously, as the strain is autotrophic nitrogen removal, no additional carbon source is needed to be added to ensure the activity of the strain; on the other hand, in the embodiment of the invention, the sterilized sewage is used as the culture medium raw material, the cost of industrial production is reduced, and the concentration of the orthogonally optimized nutrient elements is higher than that of the bacteria liquid cultured by a pure sterilized sewage culture medium, and the time for reaching the lag phase is short, so that the ammonia nitrogen removal speed of the ammonia oxidation bacteria agent obtained by the invention is high, and the total nitrogen removal efficiency is improved.
As described above, the invention reduces the problem of nutrition competition of different strains in sewage with low carbon nitrogen ratio by screening aerobic ammonia oxidizing bacteria for autotrophic denitrification, improves the overall concentration of effective bacteria, and preferably selects a culture medium taking sterilized sewage as a raw material, and adds a proper amount of nutrient elements to improve the concentration of cultured bacteria liquid, thereby improving the removal efficiency of ammoniacal nitrogen, and consequently improving the removal rate of total nitrogen.
Drawings
FIG. 1 shows a colony diagram of aerobic ammonia oxidizing bacteria N1 in an embodiment of the invention;
FIG. 2 shows a scanning electron micrograph of aerobic ammonia oxidizing bacteria N1 according to an embodiment of the present invention;
FIG. 3 shows a phylogenetic tree of aerobic ammonia oxidizing bacteria N1 according to an embodiment of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings and detailed description so that the above objects, features and advantages of the present invention can be more clearly understood. The following examples are given for the detailed implementation and specific operation of the present invention, but the scope of the present invention is not limited to the following examples.
The applicant of the invention finds that the carbon-nitrogen ratio in the sewage is reduced in recent years, the total nitrogen emission standard is difficult to reach by the traditional treatment industry,
in order to solve the problems, the invention provides an ammonia oxidizing bacteria agent which can efficiently degrade ammonia nitrogen and is practically produced and applied and a preparation method thereof.
In a first aspect, the invention provides an ammonia oxidizing bacteria agent for treating ammonia nitrogen in domestic sewage, wherein the agent comprises one or more aerobic ammonia oxidizing bacteria.
Preferably, the aerobic ammonia oxidizing bacteria are gram-positive bordetella.
Preferably, the concentration of the microbial inoculum is 108-109one/mL.
Preferably, the aerobic ammonia oxidizing bacteria have positive reactions of contact enzyme and oxidase; starch hydrolysis, gelatin liquefaction, methyl red reaction, V-P reaction, and citrate utilization reaction were all negative.
The aerobic ammonia oxidizing bacteria are autotrophic denitrogenation bacteria, and can metabolize pollutant ammonia nitrogen in the domestic sewage with low carbon-nitrogen ratio by taking the pollutant ammonia nitrogen in the sewage as an energy source, so that the dependence on a carbon source is reduced, and meanwhile, as the aerobic ammonia oxidizing bacteria belong to the aerobic ammonia oxidizing bacteria, compared with the anaerobic ammonia oxidizing bacteria and ammonia oxidizing archaea, the aerobic ammonia oxidizing bacteria in the embodiment of the invention are easier to enrich and separate, and can realize expanded culture so as to be applied to actual sewage treatment.
In a second aspect, a method for preparing an ammonia oxidizing bacteria agent for ammonia nitrogen treatment of domestic sewage comprises the following steps: inoculating one or more aerobic ammonia oxidizing strains into a culture medium, and culturing at 30 ℃ and 120r/min for 24h to obtain an ammonia oxidizing bacterium agent.
Preferably, the raw material of the culture medium is sterilized sewage, and the culture medium further comprises beef extract, trace elements, magnesium elements and potassium elements.
Preferably, the concentrations of the beef extract, the trace elements, the magnesium elements and the potassium elements in the culture medium are 5g/L, 0.5mL/L, 1.0mg/L and 1.5mg/L respectively.
The sterilized sewage is wide in source in actual sewage treatment and can be directly taken, zero cost of raw materials is realized, and in addition, the concentrations of beef extract, trace elements, magnesium elements and potassium elements which are preferably selected through an orthogonal test are respectively 5g/L, 0.5mL/L, 1.0mg/L and 1.5mg/L, so that the maximum specific growth rate of the strain in the embodiment of the invention is highest, the lag phase time is shortened, the maximum growth amount is reached, the treatment speed of ammoniacal nitrogen is greatly improved, and the removal efficiency of total nitrogen is improved.
Preferably, the one or more aerobic ammonia oxidizing strains are selected by:
step 1: standing the sludge-water mixture taken out of the aeration tank, and removing supernatant to obtain sludge;
step 2: taking part of the sludge in a centrifugal tube, diluting with sterile normal saline and fully oscillating;
and step 3: taking a preset milliliter of the oscillated liquid to a conical flask filled with a domestication culture medium, and placing the conical flask in a constant-temperature shaking table to culture in a dark place; wherein the rotating speed of the constant-temperature shaking table is a preset rotating speed;
preferably, the preset mL is 5 mL; the preset rotating speed is 120r/min
And 4, step 4: measuring the concentration of ammoniacal nitrogen in the culture solution in the constant-temperature shaking table at intervals of preset time, when the concentration of ammoniacal nitrogen is lower than a preset concentration value, taking the preset milliliter of culture solution, transferring the preset milliliter of culture solution into a fresh domestication culture medium, and repeating the step 4;
preferably, the preset time is 2 days, and the preset concentration value is 50 mg/L;
and 5: and (4) after repeating the step (4) for preset times, taking the culture solution to perform gradient dilution, flat plate coating and streak separation so as to screen out one or more aerobic ammonia oxidizing strains.
Preferably, the preset number of times is 3;
in order to make the present invention more understandable to those skilled in the art, an ammonia oxidizing bacteria agent for domestic sewage treatment and a method for preparing the same according to the present invention will be described below with reference to a plurality of specific examples.
Example 1:
preference for the aerobic ammonia oxidizing bacteria strain:
the invention takes the sludge-water mixture taken out from the aeration tank to stand for 30min, abandons the supernatant, takes 5mL to 50mL centrifugal tubes, uses sterile normal saline to dilute 10 times, and fully oscillates for 5min on a homogeneous oscillator. Then, 5mL of the medium is taken out to a conical flask filled with 150mL of the acclimatization medium, and the medium is placed in a constant temperature shaking table at 30 ℃ and 120r/min for culturing in the dark.
Measuring NH in the solution every 2 days4 +-N、NO2 --N、NO3 -Concentration of-N when NH4 +When the N concentration is lower than 50mg/L, 5mL of culture solution is taken to be transferred into a fresh domestication culture medium for continuous domestication culture, and after 3 times of transfer, the culture solution is taken for gradient dilution, plate coating and streak separation. And (3) separating the aerobic ammonia oxidizing bacteria by adopting a direct screening mode.
The separation culture medium comprises the following components: (NH)4)2SO4 0.5g/L，NaCl 2g/L，FeSO4·7H2O 0.04g/L，MgSO4·7H2O 0.5g/L，KH2PO4 3H2O 1g/L，CaCO35g/L agar 20g/L, pH adjusted to 7.2.
The invention obtains 5 aerobic ammonia oxidizing bacteria by a direct separation method, which are respectively named as N1-N5. After 48h of nitration reaction, N1 is reacted with NH4 +The removal rate of-N can reach 28.28%. Compared with the original sewage, the ammonia-oxidizing bacteria can effectively improve the ammonia nitrogen removal performance, and compared with a control group, the ammonia-oxidizing bacteria can improve the ammonia nitrogen removal effect by about 15%. Therefore, N1 is preferably a strain of an ammonia oxidizing bacterium agent.
The test characteristics of the strain N1 obtained in the example of the invention are as follows:
the colony of N1 obtained in the example of the present invention is round-point, yellowish, soft and smooth, and its colony morphology is shown in FIG. 1. The structure of N1 under a scanning electron microscope is shown in FIG. 2, and compared with other strains, the size of N1 is smaller and is close to a sphere. The aerobic ammonia oxidizing bacteria N1 are gram-positive bacteria, and the reactions of catalase and oxidase are both positive; starch hydrolysis, gelatin liquefaction, methyl red reaction, V-P reaction, and citrate utilization reaction were all negative. N1 phylogenetic tree As shown in FIG. 3, it was found that the bacterium had the highest similarity to Bordetella.
Control run 2:
the control experiment shows that the ammonia nitrogen removal effects of the aerobic ammonia oxidizing bacteria N1 obtained by direct screening and domestication screening are compared with that of the aerobic ammonia oxidizing bacteria ZX 1:
a250 mL conical flask is used, 30mL return sludge of a secondary sedimentation tank and 120mL domestic sewage are added into the conical flask, and the sludge sedimentation ratio and the suspended solid concentration of the mixed solution are respectively controlled to be about 20 percent and 1500 mg/L. Respectively inoculating the directly screened strain N1 and the domesticated and screened strain ZX1 to a beef extract peptone liquid culture medium, and fermenting and culturing for 48h until the microbial inoculum concentration is about 1010one/mL. Adding two microbial inoculum into a conical flask according to the ratio of 1:1000 of the microbial inoculum to the sewage volume, placing the conical flask in a constant temperature shaking table at 30 ℃ and 120r/min, and taking supernatant of 8h to measure NH4 +-N concentration.
Setting untreated sewage as control group, placing in constant temperature shaking table at 0 deg.C and 120r/min, collecting supernatant for 8 hr, and measuring NH4 +-N concentration.
The domestication culture medium comprises the following components: (NH)4)2SO4 0.5g/L，NaCl0.3 g/L，FeSO4·7H2O 0.03g/L，MgSO4·7H2O 0.03g/L，KH2PO4·3H2O 1g/L，NaHCO33.6g/L, pH adjusted to 7.8.
The beef extract peptone medium comprises the following components: 3g/L of beef extract, 10g/L of peptone and 5g/L of NaCl, and adjusting the pH to 7.4-7.6.
The results show that NH of the control group4 +The removal of-N was 39.51%; NH of N1 and ZX14 +the-N removal rates are 65.80% and 54.20%, respectively, and are remarkably improved compared with a control group, the NH4+ -N removal rate can be improved by 25% to the maximum extent, which indicates that the treatment effect of ammoniacal nitrogen of the obtained bacterial strain by adopting a direct screening mode is better, and the microbial inoculum obtained by the embodiment of the invention can remarkably improve the NH of sewage by adding the microbial inoculum4 +-N removal rate.
Control run 3:
this control experiment shows that the culture medium of the ammonia oxidizing bacteria agent of the present application compares with the pure sterile wastewater culture medium:
in order to enable the added strains to become dominant strains in the activated sludge and reduce the culture cost, the sterilized domestic sewage is selected as a culture medium, and beef extract, potassium element, magnesium element and trace elements are added to enrich nutrient sources lacking in the culture medium. Under the condition of single culture of sterilized sewage, the concentration of N1 is only 2.3 multiplied by 106 per mL; the concentration of N1 can reach 2.8 multiplied by 108 per mL after the nutrient substances are added, and is improved by about 2 orders of magnitude compared with the pure sterilized sewage.
Orthogonal tests are carried out on the culture medium, and the optimization result shows that the influence effects of the four influence factors are beef extract, potassium element, trace element and magnesium element from large to small. When the addition amounts of the beef extract, the trace elements, the magnesium elements and the potassium elements are respectively 5g/L, 0.5mL/L, 1.0mg/L and 1.5mg/L, the maximum specific growth rate of N1 is the highest, the lag phase time is shortened, and the maximum growth amount is reached
The results show that the concentration of N1 can reach 2.8 multiplied by 10 under the condition that the addition amounts of the beef extract, the trace elements, the magnesium element and the potassium element are respectively 5g/L, 0.5mL/L, 1.0mg/L and 1.5mg/L8The concentration of N1 in the control group using sterilized wastewater alone as the culture medium was only 2.3X 106Adding the nutrient elements of beef extract, trace elements, magnesium elements and potassium elements according to the concentrations of 5g/L, 0.5mL/L, 1.0mg/L and 1.5mg/L respectively to improve the culture concentration of N1 by 2 orders of magnitude. Meanwhile, the maximum specific growth rate of N1 is the highest, the lag phase time is shortened and the maximum growth amount is reachedCompared with the traditional culture medium, the culture medium shortens the time to 4h, which shows that in the culture medium of the embodiment, the time for the N1 to reach the stationary phase is shorter, the concentration of the microbial inoculum is higher, and the concentration of effective bacteria for treating sewage is improved, so that the removal efficiency of ammoniacal nitrogen is improved.
The application of the ammonia oxidizing bacteria agent in the actual domestic sewage treatment is described as follows:
inoculating aerobic ammonia oxidizing bacteria into sterilized sewage culture medium added with nutrient substances, and performing constant temperature shaking culture at 30 ℃ and 120r/min for 24h, wherein the concentration of the microbial inoculum is stabilized at 108-109one/mL.
Adding the microbial inoculum to a daily treatment capacity of 50m3The process of the pilot reactor and the sewage treatment equipment is A2The process is carried out by adopting a/O technology. The microbial inoculum is added according to the volume ratio of the microbial inoculum to daily treated sewage of 1: 10000 for 7 days.
The result shows that the removal rate of the total nitrogen is gradually increased from 60% to 70% and can be kept stable, the addition of the ammonia oxidizing bacteria can effectively improve the denitrification rate of the pilot plant sewage treatment equipment, and the removal rate of the total nitrogen can be stably increased by about 10%.
The application result shows that the microbial inoculum obtained by the embodiment of the invention can effectively improve the removal rate of total nitrogen, and because the concentration of the obtained bacterial strain is higher, ammonia nitrogen is converted into nitrite by ammonia oxidizing microorganisms, and then the nitrite is oxidized into nitrate by nitrite oxidizing bacteria, wherein the ammonia oxidation reaction is the rate-limiting step in the process; meanwhile, according to the embodiment of the invention, on one hand, the screened strains are convenient to screen, and meanwhile, as the autotrophic denitrification is adopted, no additional carbon source is required to be added to ensure the activity of the strains, so that the situation of competition of different strains on the carbon source under the condition of low carbon and nitrogen content is avoided; on the other hand, the microbial inoculum obtained by the culture medium provided by the invention can greatly shorten the time for reaching the stabilization period, and also improve the concentration of the bacterial liquid, and the increase of the effective bacterial concentration accelerates the sewage treatment efficiency, and simultaneously, in the practical application, after the microbial inoculum is continuously added for 7 days, compared with the condition without the microbial inoculum, the removal rate of the total nitrogen is gradually increased from 60% to 70% and can be kept stable, so that the removal effect is ensured, on the other hand, the culture medium adopts sterilized sewage as the raw material, so that the industrial production cost of the microbial inoculum is reduced, and the application of the microbial inoculum in the practical sewage treatment is facilitated.
Conventional operations in the operation steps of the present invention are well known to those skilled in the art and will not be described herein.
For simplicity of explanation, the method embodiments are described as a series of acts or combinations, but those skilled in the art will appreciate that the present invention is not limited by the order of acts, as some steps may occur in other orders or concurrently in accordance with the invention. Further, those skilled in the art will appreciate that the embodiments described in the specification are preferred embodiments and that the acts and elements referred to are not necessarily required to practice the invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The ammonia oxidizing bacteria agent applied to sewage ammonia nitrogen treatment and the preparation method thereof provided by the invention are described in detail, the principle and the implementation mode of the invention are explained by applying specific examples, and the description of the examples is only used for helping to understand the method and the core idea of the invention; meanwhile, for a person skilled in the art, according to the idea of the present invention, there may be variations in the specific embodiments and the application scope, and in summary, the content of the present specification should not be construed as a limitation to the present invention.

Claims (8)

1. An ammonia oxidizing bacteria microbial inoculum applied to sewage ammonia nitrogen treatment is characterized by comprising one or more aerobic ammonia oxidizing bacteria.
2. The ammonia oxidizing bacteria preparation of claim 1, wherein said aerobic ammonia oxidizing bacteria are gram-positive bordetella.
3. The ammonia oxidizing bacterial agent of claim 1, wherein the concentration of said agent is 108-109one/mL.
4. The ammonia oxidizing bacteria inoculant according to claim 1, wherein both the aerobic ammonia oxidizing bacteria catalase and oxidase reactions are positive; starch hydrolysis, gelatin liquefaction, methyl red reaction, V-P reaction, and citrate utilization reaction were all negative.
5. A preparation method of an ammonia oxidizing bacteria agent for treating ammonia nitrogen in domestic sewage is characterized by comprising the following steps: inoculating one or more aerobic ammonia oxidizing strains into a culture medium, and culturing at 30 ℃ and 120r/min for 24h to obtain an ammonia oxidizing bacterium agent.
6. The method for preparing an ammonia oxidizing bacterium agent according to claim 5, wherein the raw material of the culture medium is sterilized sewage, and the culture medium further comprises beef extract, trace elements, magnesium elements and potassium elements.
7. The culture medium according to claim 6, wherein the concentrations of beef extract, trace elements, magnesium elements and potassium elements in the culture medium are 5g/L, 0.5mL/L, 1.0mg/L and 1.5mg/L respectively.
8. The method of claim 5, wherein the one or more aerobic ammonia oxidizing strains are selected by:
step 1: standing the sludge-water mixture taken out of the aeration tank, and removing supernatant to obtain sludge;
step 2: taking part of the sludge in a centrifugal tube, diluting with sterile normal saline and fully oscillating;
and step 3: taking a preset milliliter of the oscillated liquid to a conical flask filled with a domestication culture medium, and placing the conical flask in a constant-temperature shaking table to culture in a dark place; wherein the rotating speed of the constant-temperature shaking table is a preset rotating speed;
and 4, step 4: measuring the concentration of ammoniacal nitrogen in the culture solution in the constant-temperature shaking table at intervals of preset time, when the concentration of ammoniacal nitrogen is lower than a preset concentration value, taking the preset milliliter of culture solution, transferring the preset milliliter of culture solution into a fresh domestication culture medium, and repeating the step 4;
and 5: and (4) after repeating the step (4) for preset times, taking the culture solution to perform gradient dilution, flat plate coating and streak separation so as to screen out one or more aerobic ammonia oxidizing strains.
CN202111425619.3A 2021-11-26 2021-11-26 Ammonia oxidizing bacteria agent applied to sewage ammonia nitrogen treatment and preparation method thereof Pending CN114231441A (en)

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CN101306903A (en) * 2007-05-18 2008-11-19 中国石油化工股份有限公司 Biochemical treatment process for high-concentration ammonia-nitrogen-containing waste water
CN101654663A (en) * 2009-03-24 2010-02-24 浙江大学 Bordetella strain and application thereof
CN101735996A (en) * 2009-12-10 2010-06-16 中国科学院地理科学与资源研究所 Mixed bactericide for restoring places polluted by polynuclear aromatic hydrocarbons, preparation method and application method

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