CN112501075A - Application of alcaligenes faecalis in preparation of microbial agent for efficiently degrading ammonia nitrogen and nitrite nitrogen in aquaculture water - Google Patents
Application of alcaligenes faecalis in preparation of microbial agent for efficiently degrading ammonia nitrogen and nitrite nitrogen in aquaculture water Download PDFInfo
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Abstract
The invention provides an application of alcaligenes faecalis in preparing a microbial agent for efficiently degrading ammonia nitrogen and nitrite nitrogen in an aquaculture water body. The microbial agent comprises the microbial content of not less than 5.0 x 108CFU/g of alcaligenes faecalis powder and a carrier, wherein the mass ratio of the alcaligenes faecalis powder to the carrier is 1: 50-100. The alcaligenes faecalis powder is a bacillus faecalis powder with the preservation number of CGMCC No.20364The strain GBW-HB1905 is prepared, has wide salt tolerance, and has the growth salinity range of 15-35 per mill. The microbial inoculum has the capability of efficiently and rapidly degrading ammonia nitrogen and nitrite nitrogen in the aquaculture water body, can continuously and stably maintain the ammonia nitrogen and the nitrite nitrogen in a lower concentration range, can effectively solve and improve the problem of high concentration of the ammonia nitrogen and the nitrite nitrogen in the aquaculture water body, and is beneficial to aquaculture and income.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and relates to application of alcaligenes faecalis in preparing a microbial agent for efficiently degrading ammonia nitrogen and nitrite nitrogen in an aquaculture water body.
Background
With the continuous improvement of the culture level in recent years, the culture density is continuously increased while higher economic benefits are pursued, so that the key is to ensure the health of the water quality of a culture water body under the condition of high-load culture density, the increase of the culture density inevitably increases the bait feeding amount of a pool body, the content of generated residual bait and undigested organic matters is also continuously increased and exceeds the self-repairing capability of the water body, particularly in prawn culture, the protein content in the bait is richer, more organic nitrogen sources are remained in the culture water body, particularly under the condition of insufficient oxygen, excessive nitrogen sources are more easily converted into toxic substances such as ammonia nitrogen, nitrite and the like, the accumulation of the toxic substances can cause great influence on the culture environment, seriously harms the health of fishes and shrimps and increases the culture risk.
The source of ammonia nitrogen is mainly the result of ammoniation of excess organic nitrogen in water by conventional heterotrophic microorganisms, namely, the organic nitrogen is decomposed into ammonia and ammonia compounds under the action of ammoniated bacteria or fungi; nitrite is mainly derived from the action of ammonia oxidizing bacteria, and can convert ammonia nitrogen into nitrite nitrogen. The harm of ammonia nitrogen to aquatic organisms is mainly from the harm of nonionic ammonia, and after the nonionic ammonia enters the aquatic organisms, the metabolic reaction and the membrane stability of the aquatic organisms are obviously influenced, so that the phenomena of difficult breathing, no ingestion, reduced resistance, convulsion, coma and the like of the cultured organisms occur, the biochemical indexes and the health conditions of the aquatic organisms are influenced, and the cultured organisms can die in batches in severe cases; nitrite poisoning can directly affect and weaken the oxygen transportation capability of cultured organisms in blood, so that the cultured organisms have the symptom of oxygen deficiency, and the appearance and internal organs of the cultured organisms are abnormal, and further the cultured organisms die. Therefore, the excessive existence of the two substances can seriously affect the health condition of aquatic organisms in the aquaculture water body, and are two large 'invisible killers' in the aquaculture water body, and if the content of the two substances is too high, the two substances possibly cause great loss to aquaculture farmers. At present, the stipulations in fishery water quality standard are as follows: during aquaculture, the concentration of ammonia should be controlled below 0.2mg/L, and should not exceed 0.6mg/L during actual aquaculture. And the total ammonia amount is required to be less than 0.5mg/L in the prawn culture process, and the ammonia nitrogen content is controlled within 0.2mg/L, so that the prawn culture method is safe. In addition, for nitrite, the content of the nitrite in aquaculture water is regulated to be below 0.2mg/L in fishery water quality standard, and the nitrite concentration in a shrimp pond in the actual aquaculture process is not more than 0.15mg/L, but in order to ensure the safety of aquaculture organisms, the nitrite in the water body is generally controlled to be below 0.1mg/L as much as possible during the aquaculture process. Therefore, seeking methods and modes for controlling and solving the high concentration of ammonia nitrogen and nitrite nitrogen in aquaculture water bodies become key factors influencing the success of aquaculture at present.
Disclosure of Invention
The invention provides an application of alcaligenes faecalis in preparing a microbial agent for efficiently degrading ammonia nitrogen and nitrite nitrogen in an aquaculture water body. The microbial agent comprises alcaligenes faecalis GBW-HB1905 bacterial powder and a carrier, and can efficiently remove ammonia nitrogen and nitrite nitrogen in aquaculture wastewater.
In order to achieve the purpose of the invention, the invention is realized by the following technical scheme:
the invention provides an application of alcaligenes faecalis in preparing a microbial agent for efficiently degrading ammonia nitrogen and nitrite nitrogen in an aquaculture water body.
Further, the microbial agent comprises the microbial content of not less than 5.0 multiplied by 108CFU/g Alcaligenes faecalis powder andand (3) a carrier.
Further, the mass ratio of the alcaligenes faecalis powder to the carrier is 1: 50-100.
Further, the alcaligenes faecalis powder is prepared from alcaligenes faecalis GBW-HB1905 with the preservation number of CGMCC No. 20364.
Further, the preparation method of the alcaligenes faecalis powder comprises the following steps:
(1) inoculating the Alcaligenes faecalis GBW-HB1905 into an improved NB culture medium, and culturing for 22h in a constant-temperature shaking table with the pH of 7.5-8.5 and the temperature of 28 ℃ to obtain an Alcaligenes faecalis GBW-HB1905 seed solution;
(2) inoculating 8-10% of the Alcaligenes faecalis GBW-HB1905 seed solution into a fermentation medium according to the volume ratio, adjusting the tank pressure to 0.1-0.2 MPa, the temperature to 28-30 ℃, the dissolved oxygen to be not less than 25%, stirring at 160-180 rpm, and fermenting for 16-20h to obtain an Alcaligenes faecalis GBW-HB1905 zymocyte solution;
(3) mixing the bacillus alcaligenes faecalis GBW-HB1905 zymocyte liquid and a carrier according to the volume-mass ratio of 1: 2-4, and carrying out vacuum freeze drying at the temperature of-40 to-50 ℃ to obtain bacillus alcaligenes faecalis GBW-HB1905 bacterial powder.
Furthermore, the Alcaligenes faecalis GBW-HB1905 has broad salt tolerance, and the growth salinity range is 15-35 per mill.
Furthermore, the growth temperature of the Alcaligenes faecalis GBW-HB1905 is 15-40 ℃, and the optimal growth temperature is 25-32 ℃.
Further, the growth pH value of the Alcaligenes faecalis GBW-HB1905 is 6-9, and the optimum pH value is 7.5-8.5.
Furthermore, the Alcaligenes faecalis GBW-HB1905 has a high propagation speed, enters a logarithmic growth phase after being cultured for 4 hours, enters a terminal logarithmic growth phase after being cultured for 14-16 hours, and the number of bacteria reaches 6.0 multiplied by 109CFU/mL。
Furthermore, the Alcaligenes faecalis GBW-HB1905 has the effect of efficiently degrading ammonia nitrogen and nitrite.
Further, suitable growth media for the Alcaligenes faecalis GBW-HB1905 include glucose, yeast powder, peptone and corn steep liquor dry powder.
Further, the carrier is at least one of calcium carbonate, talcum powder and diatomite.
Further, the use method of the microbial agent comprises the following steps: the microbial agent is directly added to a circulating water filter material box body, the pH value of the culture water body is kept to be 7.0-8.0, and the temperature is 20-30 ℃.
Furthermore, the addition concentration of the microbial inoculum in the aquaculture water body is 80-100 ppm.
Further, the microbial agent can degrade the ammonia nitrogen concentration in the aquaculture water body to be less than 0.1 mg/L; can degrade the nitrite nitrogen concentration to less than 0.02 mg/L.
Compared with the prior art, the invention has the following advantages and technical effects:
the Alcaligenes faecalis GBW-HB1905 disclosed by the invention is separated from a water body environment containing high-salinity wastewater, has a salt-tolerant growth characteristic, and can grow well within a salinity range of 15-35 per thousand. The microbial agent containing the alcaligenes faecalis GBW-HB1905 has the capability of rapidly degrading ammonia nitrogen and nitrite nitrogen in aquaculture water, can control the ammonia nitrogen concentration in the aquaculture water to be within 0.1mg/L, and the nitrite nitrogen concentration to be within 0.02mg/L, which is far lower than the national standard. Therefore, the microbial inoculum can be used for developing products for removing ammonia nitrogen and nitrite nitrogen in aquaculture water, can effectively relieve and solve the problem of high concentration accumulation of ammonia nitrogen and nitrite nitrogen in the aquaculture process when being applied to the water, reduces aquaculture risks, can further ensure the ecological health of the water and increase the aquaculture effect and income of aquatic products by adjusting the water quality, and therefore, has important application value and significance for aquaculture.
Drawings
FIG. 1 is a colony morphology of the Alcaligenes faecalis GBW-HB1905 on modified Nutrient Broth (NB) solid medium plates.
FIG. 2 is a growth curve of the Alcaligenes faecalis GBW-HB 1905.
FIG. 3 shows the addition position of the Alcaligenes faecalis GBW-HB1905 in aquaculture experiments.
FIG. 4 is a schematic diagram of the circulating aquaculture test system for Alcaligenes faecalis GBW-HB 1905.
FIG. 5 shows the influence of the Bacillus foecalis alkaligenes GBW-HB1905 microbial inoculum on the ammonia nitrogen concentration change in an aquaculture circulating water system.
FIG. 6 shows the influence of the Alcaligenes faecalis GBW-HB1905 microbial inoculum on the change of nitrite nitrogen concentration in an aquaculture circulating water system.
Detailed Description
The technical solution of the present invention will be further described in detail with reference to the following specific examples.
In the following examples, unless otherwise specified, the experimental methods used were all conventional methods, and materials, reagents and the like used were all available from biological or chemical reagents companies.
The formulations of the media required in the following examples are as follows:
1. enriching and screening a liquid culture medium by high-salinity ammonia nitrogen degrading bacteria: glucose 5g, NH4Cl 1g，K2HPO4 1.0g，MgSO4·7H2O 0.1g，FeSO4·7H20.1g of O, 30g of NaCl, 7.0-8.0 of pH and 1L of distilled water.
2. Separating and purifying the solid culture medium by the high-salinity nitrate nitrogen degrading bacteria: glucose 5g, NH4Cl 1g，K2HPO41.0g，MgSO4·7H2O 0.1g，FeSO4·7H20.1g of O, 30g of NaCl, 15g of agar powder, 7.0-8.0 of pH and 1L of distilled water.
3. Modified Nutrient Broth (NB) medium: beef extract 5g, peptone 10g, sodium chloride 5g, NH4Cl 2g/L、FeSO4·7H2O 0.1g/L、MgSO4·7H20.1g/L of O and 1L, pH 7.0.0-7.5 of water.
4. Modified Nutrient Broth (NB) solid medium: beef extract 5g, peptone 10g, sodium chloride 5g, NH4Cl2g/L、FeSO4·7H2O 0.1g/L、MgSO4·7H2O0.1 g/L, Qiongqiong15g of fat powder and 1L, pH 7.0.0-7.5 g of water.
5. Fermentation medium: 30g/L glucose, 5g/L peptone, 5g/L yeast powder and 5g/L, NaCl 10g/L, NH corn steep liquor dry powder4Cl 2g/L、KH2PO4 0.5g/L、FeSO4·7H2O 0.1g/L、Na2HPO4 2g/L、MgSO4·7H20.1g/L of O, the balance of water, and the pH value of the liquid culture medium is 7.0-7.5.
The above culture medium is sterilized at 121 deg.C for 20min before use, and then stored at room temperature.
Example 1: screening, separation and identification of Alcaligenes faecalis GBW-HB1905
1. Screening and purifying Alcaligenes faecalis GBW-HB1905
Taking 15mL of mixed sewage to be treated in a municipal sewage plant, adding the mixed sewage into 135mL of PBS buffer solution, oscillating for 20min to fully mix the samples, centrifuging for 1min at 1000rpm, and collecting the sample supernatant for later use. And adding 1mL of the sample supernatant into a conical flask filled with 100mL of the high-salinity ammoniacal nitrogen degrading bacterium enrichment screening liquid culture medium, and carrying out shaking culture at the constant temperature of 28 ℃ and 180rpm for 2d for enrichment for 3 times. After the cultured bacterial suspension is diluted in a gradient way, 100 mu L of the bacterial suspension is coated on a solid culture medium for enriching and screening the high-salinity ammonia nitrogen degrading bacteria and is placed in an incubator at 28 ℃ for culture. After 48h, single colonies of different morphologies were picked and streaked on modified Nutrient Broth (NB) solid medium, and isolation and purification were repeated 3 times to obtain a single colony, which was named as GBW-HB 1905.
As shown in figure 1, the bacterial colony of the bacterial strain GBW-HB1905 on a modified Nutrient Broth (NB) solid medium plate is round, milk white, 1-2mm in diameter, smooth, moist and slightly glossy in surface, flatter, slightly convex in middle, opaque, neat in edge and free of halo.
2. Identification of Alcaligenes faecalis GBW-HB1905
The DNA of the strain GBW-HB1905 is used as a template, 16S rRNA universal primers are used for amplification, sequence determination is carried out on amplified fragments, the 16S rDNA sequencing result of the obtained strain GBW-HB1905 is compared with the sequence in GenBank for analysis, and the result shows that the homology of the strain GBW-HB1905 and Alcaligenes faecalis is the highest, so that the GBW-HB1905 strain is determined to be Alcaligenes faecalis.
And (3) performing strain preservation on the screened strain GBW-HB1905, wherein the preservation unit of the Alcaligenes faecalis GBW-HB1905 is as follows: china general microbiological culture Collection center (CGMCC); address: western road No. 1, north west city of township, beijing, institute of microbiology, china academy of sciences; the preservation date is as follows: year 2020, month 07, 15; the preservation number of the Alcaligenes faecalis is as follows: CGMCC No. 20364.
Example 2: growth determination and physiological and biochemical characteristics of Alcaligenes faecalis GBW-HB1905
1. Growth assay for Alcaligenes faecalis GBW-HB1905
Inoculating the slant-cultured Alcaligenes faecalis GBW-HB1905 into an improved NB culture medium, culturing for 22h in a constant-temperature shaking table with the pH of 7.0-7.5 and the temperature of 28 ℃ to prepare GBW-HB1905 bacterial liquid, sampling once every 1.0h, measuring the light absorption value under OD600nm, and drawing a growth curve chart. As shown in FIG. 2, the experimental results show that GBW-HB1905 is in the growth retardation phase in the first 4h of culture, then enters the logarithmic growth phase, and enters the terminal logarithmic growth phase when being cultured for 14-16h, and the bacterial count reaches 6.0 × 109CFU/mL, 16-20h is a growth stabilization phase, followed by a decline phase, completing the entire growth cycle.
2. Physiological and biochemical characteristics of Alcaligenes faecalis GBW-HB1905
After the prepared GBW-HB1905 bacterial liquid is further cultured in an NB culture medium, the detection and analysis are carried out on the Alcaligenes faecalis GBW-HB1905 according to a strain physiological and biochemical detection method in Bergey' Manual of identification of bacteria. The results are shown in Table 1. In addition, the Alcaligenes faecalis GBW-HB1905 can normally grow at the temperature of 15-40 ℃, and the optimal growth temperature is 25-32 ℃; can grow in the pH value range of 6-9, and the optimum growth pH value is 7.5-8.5; can normally grow in the range that the dissolved oxygen content is less than 0.5mg/L or more than 2mg/L, so the alcaligenes faecalis GBW-HB1905 belongs to facultative anaerobe; in addition, the strain has the ability of producing urease, beta-glucosidase and decomposing glycerol, and can degrade power-nitrate and Simon type citrateAcid salts and semi-solid agar. In addition, the formula of a suitable nutrient medium for fermenting and culturing the Alcaligenes faecalis GBW-HB1905 is as follows: 30g/L glucose, 5g/L peptone, 5g/L yeast powder and 5g/L, NaCl 10g/L, NH corn steep liquor dry powder4Cl 2g/L、KH2PO40.5g/L、FeSO4·7H2O 0.1g/L、Na2HPO4 2g/L、MgSO4·7H20.1g/L of O, the balance of water, and the pH value of the culture medium is 7.5-8.5.
TABLE 1 physiological and biochemical characteristics of Alcaligenes faecalis GBW-HB1905
Note: < + > represents positive, and < - > represents negative.
Example 3: salinity tolerance test of Alcaligenes faecalis GBW-HB1905
In order to test the survival performance of the Alcaligenes faecalis GBW-HB1905 under high salinity conditions, the growth of the Alcaligenes faecalis GBW-HB1905 under different salinity conditions was analyzed and detected. Adjusting the amount of sodium chloride in the NB culture medium to prepare culture media with different salt concentrations, inoculating the slant-cultured Alcaligenes faecalis GBW-HB1905 into the NB culture media with different salt concentrations, culturing for 18h in a constant-temperature shaking table with the pH value of 7.0-7.5 and the temperature of 28 ℃ to obtain a culture solution of the Alcaligenes faecalis GBW-HB1905, and detecting the growth conditions of the Alcaligenes faecalis in the culture media with different salt concentrations. As shown in Table 2, GBW-HB1905 has good salt-resistant growth characteristics, and can grow well within the salinity range of 15-35 per mill.
TABLE 2 growth of Alcaligenes faecalis GBW-HB1905 in different salt concentrations
Example 4: evaluation of influence of Alcaligenes faecalis GBW-HB1905 microbial inoculum on ammonia nitrogen and nitrite nitrogen concentration in aquaculture system
1. Preparation of Alcaligenes faecalis GBW-HB1905 microbial inoculum
(1) Inoculating the slant-cultured Alcaligenes faecalis GBW-HB1905 into an improved NB culture medium, and culturing for 22h in a constant-temperature shaking table with the pH of 7.5-8.5 and the temperature of 28 ℃ to obtain an Alcaligenes faecalis GBW-HB1905 seed solution; inoculating the Alcaligenes faecalis GBW-HB1905 seed solution into a fermentation medium according to the volume ratio of 10%, adjusting the tank pressure to 0.1MPa, the temperature to 28-30 ℃, the dissolved oxygen to be not less than 25%, stirring speed to 180rpm, and fermenting for 20h to obtain the Alcaligenes faecalis GBW-HB1905 zymocyte solution.
(2) Mixing the Alcaligenes faecalis GBW-HB1905 zymocyte liquid and a carrier according to the volume-mass ratio of 1: 2-4, and carrying out vacuum freeze drying at the temperature of-40 to-50 ℃ to obtain dry bacterial powder.
(3) Adding a carrier with the mass ratio of 1: 50-100 into the dry bacterial powder, and uniformly mixing to obtain an alcaligenes faecalis GBW-HB1905 microbial inoculum; the bacteria content of the microbial inoculum is not less than 5.0 multiplied by 108CFU/g。
Wherein the carrier is calcium carbonate, talcum powder, diatomite or a mixture of the calcium carbonate, the talcum powder and the diatomite.
2. The ammonia nitrogen concentration of the water body is detected by adopting a nano reagent spectrophotometry (HJ-535-2009) and the nitrite nitrogen concentration of the water body is detected by adopting a gas phase molecular spectrometry (HJT-197-2005). The test is divided into two groups, each group is 3 repeated, wherein the CK group is a control group without adding the Bacillus foecalis alkaligenes GBW-HB1905 microbial inoculum, the S group is a test group with 100ppm Bacillus foecalis alkaligenes GBW-HB1905 microbial inoculum, wherein the adding position of the microbial inoculum is the circulating water filtering filter material box body (as shown in figure 3), and the outline of the circulating aquaculture test system is shown in figure 4. The pH of the aquaculture water was 7.3 and the temperature was 20 ℃ at the beginning of the test. Setting the whole test tracking evaluation period to be 8 days, and tracking and detecting the change of the ammonia nitrogen and nitrite nitrogen concentration in the water body at a fixed time every day. The results of the changes of the concentrations of ammonia nitrogen and nitrite nitrogen in the aquaculture water in the whole test period are respectively shown in fig. 5 and fig. 6, the concentrations of ammonia nitrogen and nitrite nitrogen in the test group are respectively reduced to 0.155mg/L and 0.085mg/L and are both within the safe concentration range by the 4 th day of the test, then the concentrations of ammonia nitrogen and nitrite nitrogen in the system are continuously reduced until the concentrations of two ammonia nitrogen and nitrite nitrogen in the system are reduced to 0.065mg/L and 0.001mg/L and are stably within the safe concentration range by the 8 th day after the test tracking is finished. The method shows that the alcaligenes faecalis GBW-HB1905 microbial inoculum can obviously reduce the concentration of ammonia nitrogen and nitrite nitrogen in the aquaculture water body, further can reduce the aquaculture risk and improve the aquaculture income of the aquatic product, and therefore, the microbial inoculum has important application value and significance.
The production process of the alcaligenes faecalis GBW-HB1905 microbial inoculum is mature and stable, the use method is simple and convenient in practical application, the alcaligenes faecalis is directly added to the surface of a circulating water filter material, the effect is obvious, and the alcaligenes faecalis has good industrialization prospect and value.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions.
Claims (10)
1. The application of the alcaligenes faecalis in preparing a microbial agent for efficiently degrading ammonia nitrogen and nitrite nitrogen in an aquaculture water body.
2. The use of claim 1, wherein said microbial agent comprises a microbial content of not less than 5.0 x 108CFU/g Alcaligenes faecalis powder and a carrier.
3. The use of claim 2, wherein the mass ratio of the alcaligenes faecalis powder to the carrier is 1: 50-100.
4. The use according to claim 2, wherein the powder of Alcaligenes faecalis is prepared from Alcaligenes faecalis GBW-HB1905 with the accession number CGMCC No. 20364.
5. The use according to claim 4, wherein the powder of Alcaligenes faecalis is prepared by the steps of:
(1) inoculating the Alcaligenes faecalis GBW-HB1905 into an improved NB culture medium for culture to obtain an Alcaligenes faecalis GBW-HB1905 seed solution;
(2) inoculating 8-10% of the Alcaligenes faecalis GBW-HB1905 seed solution into a fermentation medium according to the volume ratio, adjusting the tank pressure to 0.1-0.2 MPa, the temperature to 28-30 ℃, the dissolved oxygen to be not less than 25%, stirring at 160-180 rpm, and fermenting for 16-20h to obtain an Alcaligenes faecalis GBW-HB1905 zymocyte solution;
(3) mixing the bacillus alcaligenes faecalis GBW-HB1905 zymocyte liquid and a carrier according to the volume-mass ratio of 1: 2-4, and carrying out vacuum freeze drying at the temperature of-40 to-50 ℃ to obtain bacillus alcaligenes faecalis GBW-HB1905 bacterial powder.
6. The use of claim 4, wherein the Alcaligenes faecalis GBW-HB1905 has a salt tolerance of 15-35% per mill.
7. The use according to claim 2, wherein the carrier is at least one of calcium carbonate, talc and diatomaceous earth.
8. The use according to claim 2, wherein the microbial agent is used by: the microbial agent is directly added to a circulating water filter material box body, the pH value of the culture water body is kept to be 7.0-8.0, and the temperature is 20-30 ℃.
9. The use of claim 8, wherein the microbial agent is added to the aquaculture water at a concentration of 80-100 ppm.
10. The use of claim 1, wherein the microbial agent is capable of degrading ammonia nitrogen concentration in an aquaculture water to less than 0.1 mg/L; can degrade the nitrite nitrogen concentration to less than 0.02 mg/L.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114590911A (en) * | 2022-05-09 | 2022-06-07 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Denitrifying microbial inoculum with broad-spectrum dissolved oxygen and organic carbon tolerance and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101565240A (en) * | 2009-05-23 | 2009-10-28 | 无锡绿水之源生物科技有限公司 | Method for preparing water quality modifying microecological preparation for reducing ammonia nitrogen and nitrite nitrogen |
| CN103289939A (en) * | 2013-06-19 | 2013-09-11 | 重庆大学 | Alcaligenes faecalis and application thereof |
| WO2015046664A1 (en) * | 2013-09-27 | 2015-04-02 | 주식회사 두산에코비즈넷 | Alcaligenes faecalis ebn-ns13 (kctc 12471bp) having remarkable nitrification and denitrification capacity |
| CN108676752A (en) * | 2018-05-31 | 2018-10-19 | 中国水产科学研究院珠江水产研究所 | Bacillus foecalis alkaligenes Y311 screening techniques and application |
| CN111117912A (en) * | 2019-12-28 | 2020-05-08 | 北京翰祺环境技术有限公司 | Salt-tolerant denitrifying bacteria strain, salt-tolerant denitrifying bacteria liquid, screening method and application |
-
2020
- 2020-12-17 CN CN202011490778.7A patent/CN112501075B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101565240A (en) * | 2009-05-23 | 2009-10-28 | 无锡绿水之源生物科技有限公司 | Method for preparing water quality modifying microecological preparation for reducing ammonia nitrogen and nitrite nitrogen |
| CN103289939A (en) * | 2013-06-19 | 2013-09-11 | 重庆大学 | Alcaligenes faecalis and application thereof |
| WO2015046664A1 (en) * | 2013-09-27 | 2015-04-02 | 주식회사 두산에코비즈넷 | Alcaligenes faecalis ebn-ns13 (kctc 12471bp) having remarkable nitrification and denitrification capacity |
| CN108676752A (en) * | 2018-05-31 | 2018-10-19 | 中国水产科学研究院珠江水产研究所 | Bacillus foecalis alkaligenes Y311 screening techniques and application |
| CN111117912A (en) * | 2019-12-28 | 2020-05-08 | 北京翰祺环境技术有限公司 | Salt-tolerant denitrifying bacteria strain, salt-tolerant denitrifying bacteria liquid, screening method and application |
Non-Patent Citations (4)
| Title |
|---|
| JI XIANG YANG等: "Bioaugmentation with A. faecalis strain NR for achieving simultaneous nitrogen and organic carbon removal in a biofilm reactor", 《BIORESOURCE TECHNOLOGY》 * |
| 王淼等: "零换水条件下3种异养硝化细菌对底部充氧池塘水质和尼罗罗非鱼生长、抗氧化能力的影响", 《水产学报》 * |
| 陈均利等: "同步硝化反硝化菌（Alcaligenes faecalis WT14)养殖污水脱氮效果研究", 《农业环境科学学报》 * |
| 陈青云等: "高效异养硝化细菌Alcaligenes faecalis Ni3-1的分离及其脱氨特性研究", 《环境工程》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114590911A (en) * | 2022-05-09 | 2022-06-07 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Denitrifying microbial inoculum with broad-spectrum dissolved oxygen and organic carbon tolerance and application thereof |
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