CN112625942A - Aerobic denitrifying bacterium and application thereof - Google Patents
Aerobic denitrifying bacterium and application thereof Download PDFInfo
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- CN112625942A CN112625942A CN202011379389.7A CN202011379389A CN112625942A CN 112625942 A CN112625942 A CN 112625942A CN 202011379389 A CN202011379389 A CN 202011379389A CN 112625942 A CN112625942 A CN 112625942A
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- nitrogen
- pseudomonas
- jm22b6
- strain
- denitrification
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Abstract
The invention discloses aerobic denitrifying bacteria and application thereof. The Pseudomonas JM22B6 has a preservation number of: GDMCC No. 61212. By combining the analysis of morphology, physiology and biochemistry, 16SrRNA sequence, genome sequence and the like, the strain is a new species of pseudomonas. The strain is a novel aerobic denitrifying bacterium, can efficiently remove nitrate nitrogen, nitrite nitrogen and ammonium nitrogen in a water body, namely culture for 12 hours, and has 100% removal efficiency on the nitrite nitrogen and the ammonium nitrogen; the removal rate of nitrate nitrogen is 100 percent after 24 hours of culture. In addition, the bacterial strain 22B6 has better denitrification effect under the condition that the concentration of nitrite nitrogen is in the range of 14-280 mg/L. Therefore, the pseudomonas JM22B6 has important application potential in denitrification treatment of wastewater polluted by nitrogen such as nitrate nitrogen, nitrite nitrogen, ammonium nitrogen and the like, particularly wastewater polluted by various nitrite nitrogen with different concentrations.
Description
Technical Field
The invention belongs to the technical field of microorganisms and environmental engineering, and particularly relates to aerobic denitrifying bacteria and application thereof.
Background
With the rapid development of the domestic aquaculture industry, the high-density culture mode is more and more favored by people. However, the high-density cultivation can meet the demand of people on aquatic products, and causes serious harm to the cultivation environment and the surrounding ecological environment due to factors such as excessive feed investment, frequent use of antibacterial agents and the like. On one hand, the aquaculture water deteriorates, such as the accumulation of excessive baits, feces and secretions, so that the concentration of ammonium nitrogen and nitrite nitrogen in the water is increased, the healthy growth of aquaculture animals is damaged, the aquaculture yield is influenced, and the aquaculture cost is increased; on the other hand, the untreated breeding wastewater is discharged randomly, and serious pollution is caused to the water environment around the breeding. Therefore, how to remove excessive inorganic nitrogen in the aquaculture wastewater efficiently and economically has important practical significance.
The method mainly comprises three methods of physics, chemistry and biology, wherein the microorganism denitrification technology overcomes the defects of the physics and chemistry denitrification technology, and has the advantages of simple operation, wide application range, good treatment effect, no secondary pollution and the like. Therefore, the microbial denitrification technology becomes the most widely applied denitrification technology at home and abroad at present. At present, Anoxic-Aerobic (AO) denitrification, Anaerobic-Anoxic-aerobic (AAO) denitrification, Sequencing Batch Reactor (SBR) denitrification, Biological Aerated Filter (BAF) denitrification and other processes are mainly adopted in the fields of environmental engineering, sewage and wastewater treatment, and the denitrification processes are generally carried out in stages or in different reactors.
In recent years, with intensive research on denitrifying microorganisms, researchers have found that certain strains can undergo denitrification under aerobic conditions, and such strains are called aerobic denitrifying bacteria. Most Aerobic denitrifying Bacteria exhibit Heterotrophic Nitrification, and such Bacteria that have both Heterotrophic Nitrification and Denitrification functions are called Heterotrophic Nitrification-Aerobic denitrifying Bacteria (HN-AD). In view of the fact that the dissolved oxygen of the aquaculture water needs to be maintained in a certain range in the aquaculture process to ensure the healthy growth of the cultured animals, the aerobic denitrification denitrobacteria have good application value in the denitrification of the aquaculture wastewater.
The aerobic denitrifying bacteria have diversity in physiology, biochemistry and phylogeny, and are reported in documents to be mainly distributed in the genera of pseudomonas (pseudomonas), Alcaligenes (Alcaligenes), Paracoccus (Paracoccus), Bacillus (Bacillus) and the like, and related countries have more patents.
Disclosure of Invention
The first purpose of the invention is to provide an aerobic denitrifying bacterium.
The second purpose of the invention is to provide a living bacterial preparation with denitrification function.
The third purpose of the invention is to provide the application of the aerobic denitrifying bacteria or the viable bacteria preparation with denitrification function.
The purpose of the invention is realized by the following technical scheme:
an aerobic denitrifying bacterium, designated as Pseudomonas sp JM22B6, which is deposited at the Guangdong province microbial culture Collection (GDMCC) of No. 59 building of Zhou Lu 100, Pielio, Calif., Guangzhou, Guangdong province on 20 days 9-2020, with the deposit numbers: GDMCC No. 61212.
The pseudomonas JM22B6 is obtained by separating and screening water of a bighead carp and south America white prawn mixed aquaculture pond, and has the following characteristics:
1. morphological characteristics: pseudomonas JM22B6, gram-negative bacteria, NA culture medium at 30 deg.C for 48h, the colony is beige, irregular edge, slightly convex, smooth surface, and opaque, and the diameter of the colony is 2-3 mm. The cells are long rod-shaped, have the size of (0.3 multiplied by 2.4) mu m and have single extremely-growing flagella;
2. physiological and biochemical characteristics: the concentration of the growth salt of the pseudomonas JM22B6 is 0-8.0%, the pseudomonas JM22B6 can not grow normally when the concentration is more than 9%, and the optimal salt concentration is 0-2.0%; the growth pH is 6.0-11.0, most preferably 7.5; the growth temperature is 15-45 deg.C, most preferably 28-37 deg.C. Oxidase and catalase are both positive, and amylase, protease and cellulase are all negative. API 20NE results show that pseudomonas JM22B6 is positive in beta-glucosidase, arginine double hydrolase, indole production and nitrate reductase, beta-galactosidase, gelatin liquefaction and urease are negative, the strain cannot perform D-glucose fermentation to produce acid, D-glucose, D-mannitol, gluconate, capric acid, malic acid and sodium citrate can be utilized, and N-acetyl-glucosamine, D-maltose, adipic acid, L-arabinose, D-mannose and phenylacetic acid cannot be utilized. API ZYM results showed that Pseudomonas JM22B6 has enzyme activities such AS alkaline phosphatase, acid phosphatase, esterase (C4), lipid esterase (C8), lipase (C14), arylamine leucine and naphthol-AS-BI-phosphohydrolase, and does not have enzyme activities such AS α -galactosidase, β -uronidase, α -glucosidase, β -glucosidase, N-acetyl-glucosidase, α -mannosidase, β -fucosidase, arylamine valine, arylamine cystine, trypsin and chymotrypsin;
3.16S rRNA and genomic sequence analysis: the method comprises the steps of extracting genome DNA of pseudomonas JM22B6 by using a HiPure bacterial DNA extraction kit, carrying out amplification determination on a 16S rRNA sequence of the genome DNA, and sending the genome DNA to Meiji biological medicine science and technology Limited, Shanghai for whole genome sequencing, wherein the 16S rRNA sequence of a strain 22B6 is shown as SEQ ID NO. 1. 16S rRNA of Pseudomonas JM22B6 was compared with Pseudomonas mendocina NBRC14162 by BLASTTThe highest similarity was 98.5%. The strain JM22B6 and the P.mendocina NBRC14162 with the highest sequence similarity of 16S rRNA are obtainedTComparative genomic analysis showed that the average nucleotide similarity (ANI) and DNA-DNA hybridization value (dDDH) of the two strains were 84.6% and 29.0%, respectively;
according to the analysis results of morphological characteristics, physiological and biochemical characteristics, 16S rRNA sequences and genome sequences of the strain, the strain JM22B6 belongs to a new species of Pseudomonas, and particularly comprises Pseudomonas sp JM22B 6.
A viable bacteria preparation with denitrification function contains the above Pseudomonas sp JM22B 6. Pseudomonas JM22B6 as active ingredient.
The application of the aerobic denitrifying bacteria or the viable bacteria preparation with the denitrification function in the denitrification treatment of the water body.
The nitrogen includes, but is not limited to, nitrate nitrogen, nitrite nitrogen, or ammonium nitrogen. The pseudomonas JM22B6 of the strain can rapidly and efficiently remove nitrate nitrogen, nitrite nitrogen and ammonium nitrogen in a water body, namely the removal efficiency of the strain on nitrite nitrogen and ammonium nitrogen reaches 100% when the strain is cultured for 12 hours; when the strain is cultured for 24 hours, the removal rate of the nitrate nitrogen by the strain reaches 100 percent.
The denitrification mode is aerobic denitrification.
The water body can be wastewater with any over-standard nitrogen content, and the concentration of nitrite nitrogen is 14-280 mg/L. The strain Pseudomonas JM22B6 has nitrite nitrogen removal rate of over 80.0% under the condition that the concentration of nitrite nitrogen is 14-280 mg/L. Therefore, the pseudomonas JM22B6 has good denitrification effect in water bodies polluted by nitrite nitrogen with low concentration and high concentration, namely, the strain can be applied to denitrification treatment of water bodies polluted by different nitrite nitrogen concentrations.
Drawings
Figure 1 is a colony morphology map of Pseudomonas sp.jm22b 6.
FIG. 2 is a cell morphology map under transmission electron microscope of Pseudomonas sp.JM22B6.
Figure 3 is a graph of denitrification performance results of Pseudomonas sp.jm22b6 under the condition of taking nitrate nitrogen as a unique nitrogen source.
Figure 4 is a graph of denitrification performance results of Pseudomonas sp.jm22b6 under the condition of using nitrite nitrogen as a unique nitrogen source.
Figure 5 is a graph of denitrification performance results of Pseudomonas sp.jm22b6 under the condition of using ammonium nitrogen as a unique nitrogen source.
FIG. 6 is a graph showing denitrification effect of Pseudomonas sp.JM22B6 under different nitrite concentration conditions.
Detailed Description
The following examples are further illustrative of the present invention but are not intended to be limiting thereof.
Example 1: isolation, purification and preservation of Pseudomonas JM22B6
Samples were collected from the pond water of mixed culture of bighead carp and penaeus vannamei boone in Guanghai town culture base (N21 degrees 56 '31; E112 degrees 46' 16 ″) of Taishan city of Jiangmen, Guangdong province. 1mL of culture water sample and 10 percent of gradient dilution-1，10-2，10-3And 10-4Then, 200. mu.L of 10 was taken-2、10-3And 10-4The diluted solution is coated on a selective culture medium used for separating and screening denitrifying bacteria, and cultured in an incubator at 30 ℃. Visually observing colony morphology, selecting single colonies with colony morphology difference, streaking and purifying respectively, inoculating the purified colonies into 5mL of R2A liquid culture collection, culturing at 30 ℃ and 180rpm, and performing subsequent glycerol tube preservation to obtain the strain JM22B 6.
The formulation of the selective medium is as follows: sodium succinate 0.025g, sodium citrate dihydrate 0.025g, NaNO20.0069g，KNO3 0.0101g，(NH4)2SO4 0.0066g，Na2HPO4 1.0g，KH2PO4 1.0g，MgSO4·7H20.2g of O, 15g of agar, pH 7.4, constant volume of 1000mL, 121 ℃, 15min, high temperature and high pressure sterilization. When the plate is manufactured, a composite carbon source with the volume ratio of 1 percent and a trace element mixed solution with the volume ratio of 0.2 percent are added.
A composite carbon source: 13.8g of D-glucose, 13.8g of D-fructose, 13.8g of D-lactose, 12.8mL of 90% lactic acid, 14.0g of mannitol, 14.0mL of absolute ethyl alcohol, 12.6mL of glycerol, 9.6g of sodium benzoate, 9.2g of salicylic acid and 19.0g of anhydrous sodium acetate, dissolving in 1000mL of water, having pH of 7.4, and filtering and sterilizing by a 0.22-micron filter membrane.
And (3) mixing trace element liquid: EDTA-Na 58.0g, ZnSO4·7H2O 3.9g，CaCl2 10.0g，MnCl2·4H2O1.0g，FeSO4·7H2O 10.0g，(NH4)6Mo7O24·4H2O 1.1g，CuSO4·5H2O 1.6g，CoCl2·6H2O1.6 g, pH adjusted to 6.0, volume to 1000mL, 0.22 μm filter membrane filtration.
Example 2: physiological and biochemical characteristics of pseudomonas JM22B6
As shown in FIG. 1 and FIG. 2, the strain JM22B6 is a gram-negative bacterium, and when the strain is cultured on an NA medium for 48 hours, the colony morphology characteristics are as follows: beige, irregular edge, slight bulge, smooth surface and non-transparency. The cell morphology characteristics of the material observed by a transmission electron microscope are as follows: the size of the flagellum is (0.3 multiplied by 2.4) mu m, the flagellum is long-rod-shaped and has single extremely-growing flagellum.
The strain JM22B6 was inoculated into NB medium containing NaCl at 0, 0.5%, 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0% and 10.0% by mass, respectively, and cultured at 30 ℃ and 180 rpm. Strain JM22B6 was inoculated into NB medium of pH 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0 and 11.0, respectively, and cultured at 30 ℃ and 180 rpm. The strain JM22B6 was inoculated to the NA medium and cultured in incubators at 10 ℃, 15 ℃, 20 ℃, 25 ℃, 30 ℃, 35 ℃, 40 ℃, 45 ℃ and 50 ℃ respectively.
The test result shows that the pseudomonas JM22B6 can not normally grow when the salt concentration is 0-8.0 percent, the pseudomonas JM22B6 can not normally grow when the salt concentration is more than 9 percent, the optimal salt concentration is 0-2.0 percent, the growth pH is 6.0-11.0, and the optimal salt concentration is 7.5. The growth temperature is 15-45 deg.C, most preferably 28-37 deg.C.
API 20NE results show that pseudomonas JM22B6 is positive in beta-glucosidase, arginine double hydrolase, indole production and nitrate reductase, beta-galactosidase, gelatin liquefaction and urease are negative, the strain cannot perform D-glucose fermentation to produce acid, D-glucose, D-mannitol, gluconate, capric acid, malic acid and sodium citrate can be utilized, and N-acetyl-glucosamine, D-maltose, adipic acid, L-arabinose, D-mannose and phenylacetic acid cannot be utilized. The API ZYM results showed that Pseudomonas JM22B6 has enzyme activities such AS alkaline phosphatase, acid phosphatase, esterase (C4), lipid esterase (C8), lipase (C14), arylamine leucine and naphthol-AS-BI-phosphohydrolase, and does not have enzyme activities such AS alpha-galactosidase, beta-uronidase, alpha-glucosidase, beta-glucosidase, N-acetyl-glucosidase, alpha-mannosidase, beta-fucosidase, arylamine valine, arylamine cystine, trypsin and chymotrypsin. In addition, the oxidase and catalase of the strain are positive, and the amylase, protease and cellulase are negative.
Example 3: 16S rRNA sequence analysis of Pseudomonas JM22B6
Extracting genome DNA of the strain JM22B6 by using a HiPure bacterial DNA extraction kit (Guangzhou Meiji Biotechnology Co., Ltd.), amplifying a PCR product by using bacterial 16S rRNA gene amplification universal primers 27F/1492R (27F: 5 ' -AGAGT TTGATCATGGCTCAG-3, 1492R: 5'-TACGGTTACCTTGTTACGACTT-3'), sending the PCR product to Suzhou Jinzhi Biotechnology Co., Ltd for sequence sequencing, wherein the sequence is shown as SEQ ID NO.1, and performing homology comparison analysis on a sequencing result and a 16S rRNA sequence in an EzBioCloud website database to show that the strain JM22B6 and the strain Pseudomonas mendocina NBRC 62TThe similarity of the 16S rRNA gene sequence is the highest, and is 98.5 percent, which is less than the threshold value of the bacterial strain level, and is 98.7 percent. Based on the above analysis of results, the strain JM22B6 of the present invention was preliminarily identified as a novel species of Pseudomonas sp.
Example 4: genomic sequence analysis of Pseudomonas JM22B6
Extracting the genome DNA of the strain JM22B6, sending the genome DNA to Uygur biomedical science and technology Limited company for genome sequencing, and respectively adopting SPAdes v3.11.1 and CheckM 1.0.9 for genome assembly and quality evaluation according to the sequencing result. Calculation of JM22B6 and P.mendocina NBRC14162 Using the ANI Call promoter on the EzBioCloud websiteTAverage nucleotide similarity of genome (ANI); calculating JM2 by using Genome-to-Genome Distance Calculator on DSMZ website2B6 and pTDNA-DNA hybridization value (dDDH) of (1). The results showed that the strains JM22B6 and P.mendocina NBRC14162TThe ANI and dDDH values were 84.6% and 29.0%, respectively, which were less than the ANI threshold of 95% and the dDDH threshold of 70% of the bacterial species level. This result further confirmed that the strain JM22B6 is a novel species of genus Pseudomonas.
In summary, the strain JM22B6 of the present invention is a new species of Pseudomonas, named Pseudomonas sp JM22B6, which was deposited at the guangddong province collection of microorganisms (GDMCC) at 20/9/2020, address: building 59 of first furious Zhongluo 100 institute of overseas, overseas district, Guangdong province, with the preservation number: GDMCC No. 61212.
Example 5: denitrification Properties of Pseudomonas JM22B6
Inoculating strain JM22B6 into culture medium containing nitrate nitrogen, nitrite nitrogen or ammonium nitrogen as unique nitrogen source at an inoculation amount of 1%, wherein NO is3 -Initial concentration of-N set to 140mg/L, NO2 --N and NH4 +The initial concentrations of-N were all set at 50mg/L, incubated at 30 ℃ and 120 rpm. In nitrate nitrogen culture medium, sampling was performed at 0, 6, 18, 24, 30, 42, and 48h of culture, and a part of the culture solution was used for OD detection600The other part of the culture broth is centrifuged to take the supernatant for subsequent NO3 --determination of the N concentration. In nitrite nitrogen and ammonium nitrogen culture medium, sampling is carried out at 0, 12, 18, 24, 36h of culture, and a part of culture solution is used for detecting OD600The other part of the culture broth was centrifuged to obtain the supernatant, which was used for subsequent NO2 --N and NH4 +-determination of the N concentration. NO3 -The concentration of-N is determined by UV spectrophotometry, NO2 --N is determined spectrophotometrically by N- (1-naphthyl) -ethylenediamine, NH4 +The concentration of-N was determined spectrophotometrically using a Naeseler reagent.
Culture medium with nitrate nitrogen as unique nitrogen source: sodium succinate 6.08g, KNO3 1.01g，Na2HPO4 1.0g，KH2PO41.0g，MgSO4·7H2O0.2 g, addition volumeAnd (3) sterilizing the mixed solution of the trace elements with the concentration of 0.2 percent at the high temperature and the high pressure at the constant volume of 1000mL and the temperature of 115 ℃ for 20min at the pH value of 7.4.
A culture medium with nitrite nitrogen as a unique nitrogen source: sodium succinate 2.5g, trisodium citrate dihydrate 2.5g, NaNO20.24g，Na2HPO4 1.0g，KH2PO4 1.0g，MgSO4·7H20.2g of O, adding a mixed solution of a composite carbon source with the volume ratio of 1% and trace elements with the volume ratio of 0.2%, adjusting the pH to 7.4, diluting to 1000mL, adjusting the volume to 115 ℃, performing 20min, and sterilizing at high temperature and high pressure.
Medium with ammonium nitrogen as sole nitrogen source: sodium succinate 2.5g, trisodium citrate dihydrate 2.5g, (NH)4)2SO40.23g，Na2HPO4 1.0g，KH2PO4 1.0g，MgSO4·7H20.2g of O, adding a mixed solution of a composite carbon source with the volume ratio of 1% and trace elements with the volume ratio of 0.2%, adjusting the pH to 7.4, diluting to 1000mL, adjusting the volume to 115 ℃, performing 20min, and sterilizing at high temperature and high pressure.
As shown in FIG. 3, the strain JM22B6 shows that the initial concentration of NO in water is 140mg/L3 -the-N has good removing effect, namely the OD of the strain is obtained when the strain is cultured for 24 hours600Is 1.0, to NO3 -The removal effect of-N reaches 100%. As shown in FIGS. 4 and 5, the strain JM22B6 is directed to NO at an initial concentration of 50mg/L in water2 --N and NH4 +the-N can be removed rapidly and efficiently, namely the OD of the strain is 12h after the strain is cultured600Respectively 0.6 and 0.4, and the removal rate of the inorganic nitrogen reaches 100 percent. The results show that the pseudomonas JM22B6 is a novel aerobic denitrifying bacterium with a high-efficiency denitrification function, and has great application potential in denitrification of water bodies.
Example 6: denitrogenation effect of pseudomonas JM22B6 under different nitrite state concentration conditions
Inoculating strain JM22B6 into culture medium containing sodium nitrite as unique nitrogen source at an inoculation amount of 1%, respectively, and adding NO2 -Initial N concentrations were set at 3.5, 7.0, 14, 42, 70, 84, 140, 168, 210 and 280mg/L, respectively, while adjusting with sodium succinate as sole carbon sourceMaintaining the C/N in the culture medium at 15 deg.C, culturing at 30 deg.C and 120rpm for 48h, sampling, and determining the OD of the bacterial liquid600Centrifuging part of the culture solution to obtain supernatant, and measuring NO by N- (1-naphthyl) -ethylenediamine spectrophotometry2 --determination of the N concentration.
As shown in FIG. 6, strain JM22B6 is NO2 -Under the condition that the initial concentration of N is 14-280mg/L, the removal rate of nitrite nitrogen is above 80.0%, thus the strain JM22B6 can grow and denitrify under the condition of nitrite nitrogen concentration with wider concentration range, and has great application potential in the denitrification treatment of wastewater polluted by low-concentration and high-concentration nitrite nitrogen.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Sequence listing
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Claims (7)
1. An aerobic denitrifying bacterium, which is characterized in that: designated as Pseudomonas (Pseudomonas sp.) JM22B6, which was deposited at 20/9/2020 in the collection of microbial strains in Guangdong province, located at building 59 of the pioneer middle Lou 100, Uighur, Calif., Guangdong province, with the collection numbers: GDMCC No. 61212.
2. A viable bacteria preparation with denitrification function is characterized in that: contains Pseudomonas sp JM22B6 as the microorganism of claim 1.
3. The use of the aerobic denitrifying bacteria of claim 1 or the viable bacteria preparation with denitrification function of claim 2 in denitrification treatment of water body.
4. Use according to claim 3, characterized in that: the nitrogen includes, but is not limited to, nitrate nitrogen, nitrite nitrogen, or ammonium nitrogen.
5. Use according to claim 3 or 4, characterized in that: the denitrification mode is aerobic denitrification.
6. Use according to claim 3 or 4, characterized in that: the water body is waste water with excessive nitrogen content.
7. Use according to claim 3 or 4, characterized in that: the concentration of nitrite nitrogen in the water body is 14-280 mg/L.
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| CN114538704A (en) * | 2022-02-17 | 2022-05-27 | 广州昭合环保科技有限公司 | Water treatment facilities based on synchronous nitrification and denitrification technique |
| CN112625942B (en) * | 2020-12-01 | 2022-06-14 | 华南理工大学 | Aerobic denitrifying bacterium and application thereof |
| CN114874943A (en) * | 2021-12-22 | 2022-08-09 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Efficient denitrification composite preparation and application thereof |
| CN115948288A (en) * | 2022-12-06 | 2023-04-11 | 浙江大学 | Aerobic efficient denitrification compound flora and application thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112625942B (en) * | 2020-12-01 | 2022-06-14 | 华南理工大学 | Aerobic denitrifying bacterium and application thereof |
| CN114874943A (en) * | 2021-12-22 | 2022-08-09 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Efficient denitrification composite preparation and application thereof |
| CN114874943B (en) * | 2021-12-22 | 2023-10-24 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Efficient denitrification compound preparation and application thereof |
| CN114538704A (en) * | 2022-02-17 | 2022-05-27 | 广州昭合环保科技有限公司 | Water treatment facilities based on synchronous nitrification and denitrification technique |
| CN115948288A (en) * | 2022-12-06 | 2023-04-11 | 浙江大学 | Aerobic efficient denitrification compound flora and application thereof |
| CN115948288B (en) * | 2022-12-06 | 2023-09-26 | 浙江大学 | Aerobic high-efficiency denitrification compound flora and application thereof |
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